Melanocortin receptor agonists

ABSTRACT

The present invention relates to melanocortin receptor agonists of formula (I), which is useful in the treatment of obesity, diabetes and male and/or female sexual dysfunction.

FIELD OF THE INVENTION

[0001] The present invention relates to melanocortin receptor agonists,and more particularly piperazine derivatives as melanocortin receptoragonists, which are useful for the treatment or prevention of diseasesand disorders responsive to the activation of melanocortin receptors.

BACKGROUND OF THE INVENTION

[0002] Pro-opiomelanocortin (POMC) derived peptides are known to affectfood intake. Several lines of evidence support the notion that theG-protein coupled receptors (GPCRs) of the melanocortin receptor (MC-R)family, several of which are expressed in the brain, are targets of POMCderived peptides involved in the control of food intake and metabolism.

[0003] Evidence for the involvement of MC-R in obesity includes: i) theagouti (A^(VY)) mouse which ectopically expresses an antagonist of theMC-1R, MC-3R and MC-4R is obese, indicating that blocking the action ofthese three MC-Rs can lead to hyperphagia and metabolic disorders; ii)MC-4R knockout mice (Huszar et al., Cell, 88:131-141, 1997) recapitulatethe phenotype of the agouti mouse and these mice are obese; iii) thecyclic heptapeptide MC-1R, MC-3R, MC-4R, and MC-5R agonistmelanotanin-II (MT-II) injected intracerebroventricularly (ICV) inrodents, reduces food intake in several animal feeding models (NPY,ob/ob, agouti, fasted) while ICV injected SHU-9119 (MC-3R, MC-4Rantagonist; MC-1R and MC-5R agonist) reverses this effect and can inducehyperphagia; and

[0004] iv) chronic intraperitoneal treatment of Zucker fatty rats withan α-NDP-MASH derivative (HP228) has been reported to activate MC-1R,MC-3R, MC-4R and MC-5R and to attenuate food intake and body weight gainover a 12 week period.

[0005] Five MC-Rs have thus far been identified, and these are expressedin different tissues. MC-1R was initially characterized by dominant gainof function mutations at the extension locus, affecting coat color bycontrolling phaeomelanin to eumelanin conversion through control oftyrosinase. MC-1R is mainly expressed in melanocytes. MC-2R is expressedin the adrenal gland and represents the ACTH receptor. MC-3R isexpressed in the brain, gut and placenta and may be involved in thecontrol of food intake and thermogenesis. MC-4R is uniquely expressed inthe brain and its inactivation was shown to cause obesity. (A. Kask, etal., “Selective antagonist for the melanocortin-4-receptor (HS014)increases food intake in free-feeding rats, Biochem. Biophys. Res.Commun., 245:90-93, 1998). MC-5R is expressed in many tissues includingwhite fat, placenta and exocrine glands. A low level of expression isalso observed in the brain. MC-5R knock out mice reveal reducedsebaceous gland lipid production (Chen et al., Cell, 91:789-798, 1997).

[0006] MC-4R appears to play a role in other physiological functions aswell, namely controlling grooming behavior, erection and blood pressure.Erectile dysfunction denotes the medical condition of inability toachieve penile erection sufficient for successful intercourse. The term“impotence” is often times employed to describe this prevalentcondition. Synthetic melanocortin receptor agonists have been found toinitiate erections in men with psychogenic erectile dysfunction (H.Wessells et al., “Synthetic Melanotropic Petide Initiates Erections inMen With Psychogenic Erectile Dysfunction: Double-Blind, PlaceboControlled Crossover Study,” J. Urol., 160: 389-393, 1998). Activationof melanocortin receptors of the brain appears to cause normalstimulation of sexual arousal. Evidence for the involvement of MC-R inmale and/or female sexual dysfunction is detailed in WO 00/74679.

[0007] Diabetes is a disease in which a mammal's ability to regulateglucose levels in the blood is impaired because the mammal has a reducedability to convert glucose to glycogen for storage in muscle and livercells. In Type I diabetes, this reduced ability to store glucose iscaused by reduced insulin production. “Type II Diabetes” or “non-insulindependent diabetes mellitus” (NIDDM) is the form of diabetes, which isdue to a profound resistance to insulin stimulating or regulatory effecton glucose and lipid metabolism in the main insulin-sensitive tissues,muscle, liver and adipose tissue. This resistance to insulinresponsiveness results in insufficient insulin activation of glucoseuptake, oxidation and storage in muscle and inadequate insulinrepression of lipolysis in adipose tissue and of glucose production andsecretion in liver. When these cells become desensitized to insulin, thebody tries to compensate by producing abnormally high levels of insulinand hyperinsulemia results. Hyperinsulemia is associated withhypertension and elevated body weight. Since insulin is involved inpromoting the cellular uptake of glucose, amino acids and triglyceridesfrom the blood by insulin sensitive cells, insulin insensitivity canresult in elevated levels of triglycerides and LDL which are riskfactors in cardiovascular diseases. The constellation of symptoms whichinclude hyperinsulemia combined with hypertension, elevated body weight,elevated triglycerides and elevated LDL is known as Syndrome X.

[0008] Spiropiperidine and piperidine derivates have been disclosed inU.S. Pat. No. 6,294,534 B1, WO 01/70337, WO 00/74679 and WO 01/70708 asagonists of melanocortin receptor(s), which can be used for thetreatment of diseases and disorders, such as obesity, diabetes andsexual dysfunction.

[0009] In view of the unresolved deficiencies in treatment of variousdiseases and disorders as discussed above, it is an object of thepresent invention to provide novel piperazine derivatives, which areuseful as melanocortin receptor agonists to treat obesity, diabetes, andmale and female sexual dysfunction.

BRIEF SUMMARY OF THE INVENTION

[0010] The present invention relates to a compound of novel piperazinederivatives as melanocortin receptor agonists as shown formula I:

[0011] or a pharmaceutically acceptable salts or stereoisomers thereof,wherein

[0012] L and L¹ are independently: hydrogen or together oxo;

[0013] T is:

[0014] R is independently:

[0015] hydrogen,

[0016] hydroxy,

[0017] cyano,

[0018] nitro,

[0019] halo,

[0020] C₁-C₈ alkyl,

[0021] C₁-C₈ alkoxy,

[0022] C₁-C₄ haloalkyl,

[0023] (D)C(O)R⁹,

[0024] (D)C(O)OR⁹,

[0025] (D)C(O)SR⁹,

[0026] (D)C(O)heteroaryl,

[0027] (D)C(O)heterocyclyl,

[0028] (D)C(O)N(R⁹)₂,

[0029] (D)N(R⁹)₂,

[0030] (D)NR⁹COR⁹,

[0031] (D)NR⁹CON(R⁹)₂,

[0032] (D)NR⁹C(O)OR⁹,

[0033] (D)NR⁹C(R⁹)═N(R⁹),

[0034] (D)NR⁹C(═NR⁹)N(R⁹)₂,

[0035] (D)NR⁹SO₂R⁹,

[0036] (D)NR⁹SO₂N(R⁹)₂,

[0037] (D)NR⁹(CH₂)_(n)heterocyclyl,

[0038] (D)NR⁹(CH₂)_(n)heteroaryl,

[0039] (D)OR⁹,

[0040] OSO₂R⁹,

[0041] (D)[O]_(q)(C₃-C₇ cycloalkyl),

[0042] (D)[O]_(q)(CH₂)_(n)aryl,

[0043] (D)[O]_(q)(CH₂)_(n)heteroaryl,

[0044] (D)[O]_(q)(CH₂)_(n) heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen when q=1,

[0045] (D)SR⁹,

[0046] (D)SOR⁹,

[0047] (D)SO₂R⁹, or

[0048] (D)SO₂N(R⁹)₂;

[0049] wherein C₁-C₈ alkyl, C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl,heterocyclyl and heteroaryl are optionally substituted with one to fivesubstituents independently selected from R⁸;

[0050] R¹ is independently:

[0051] hydrogen, CONH(C₁-C₈ alkyl), C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇cycloalkyl or oxo, provided that oxo is not attached to the same carbonthat attached to nitrogen which forms an amide bond;

[0052] R³ is independently: aryl or thienyl;

[0053] wherein aryl and thienyl are optionally substituted with one tothree substituents selected from the group consisting of:

[0054] cyano, halo, C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy,C₁-C₄ haloalkyl and C₁-C₄ haloalkyloxy;

[0055] R⁴ is independently:

[0056] hydrogen, C₁-C₈ alkyl, C(O)R⁹, C(O)OR⁹, C₃-C₇ cycloalkyl or(CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;

[0057] each R⁸ is independently:

[0058] hydrogen,

[0059] halo,

[0060] oxo

[0061] N(R¹⁰)₂

[0062] C₁-C₈ alkyl,

[0063] (D)C₃-C₇ cycloalkyl,

[0064] C₁-C₄ haloalkyl,

[0065] C₁-C₄ alkoxy,

[0066] heteroaryl,

[0067] hydroxy,

[0068] heterocyclyl, wherein heterocyclyl excludes a heterocyclylcontaining a single

[0069] nitrogen,

[0070] phenyl,

[0071] (D)COR⁹,

[0072] (D)C(O)OR⁹

[0073] (D)OR⁹,

[0074] (D)OCOR⁹,

[0075] (D)OCO₂R⁹,

[0076] (D)SR⁹,

[0077] (D)SOR⁹, or

[0078] (D)SO₂R⁹;

[0079] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0080] each R⁹ is independently:

[0081] hydrogen,

[0082] C₁-C₈ alkyl,

[0083] C₁-C₄ haloalkyl,

[0084] (D)C₃-C₇ cycloalkyl,

[0085] (D)aryl, wherein aryl being phenyl or naphthyl,

[0086] (D)heteroaryl or

[0087] (D)heterocyclyl; wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen; and

[0088] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0089] each R¹⁰ is independently:

[0090] hydrogen, (C₁-C₈)alkyl, C(O)C₁-C₈ alkyl, aryl or C₃-C₇cycloalkyl;

[0091] each R¹¹ is independently:

[0092] hydrogen,

[0093] C₁-C₈ alkyl,

[0094] (D)aryl,

[0095] (D)heteroaryl

[0096] (CH₂)_(n)N(R⁸)₂,

[0097] (CH₂)_(n)NR⁸C(O)C₁-C₄ alkyl,

[0098] (CH₂)_(n)NR⁸SO₂C₁-C₄ alkyl,

[0099] (CH₂)_(n)SO₂N(R⁸)₂,

[0100] (CH₂)_(n)[O]_(q)C₁-C₈ alkyl,

[0101] (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸COR⁸,

[0102] (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸SO₂R⁸,

[0103] (CH₂)_(n)[O]_(q)-heterocyclyl or

[0104] (CH₂)_(n)[O]_(q)(C₁-C₈ alkyl)-heterocyclyl; and

[0105] wherein n is 2-8;

[0106] each R¹² is independently:

[0107] hydrogen,

[0108] C₁-C₈ alkyl,

[0109] (D)phenyl

[0110] C(O)C₁-C₈ alkyl,

[0111] C(O)phenyl,

[0112] SO₂C₁-C₈ alkyl or

[0113] SO₂-phenyl;

[0114] D is a bond or —(CH₂)_(n)—;

[0115] n is 0-8;

[0116] p is 0-5;

[0117] q is 0-1; and

[0118] r is 1-2.

[0119] The compounds of the present invention are useful in preventingor treating obesity or diabetes mellitus in a mammal comprising theadministration of a therapeutically effective amount of the compound offormula I.

[0120] The compounds of the present invention are also useful inpreventing or treating male or female sexual dysfunction in mammal, morespecifically erectile dysfunction, comprising the administration of atherapeutically effective amount of the compound of formula I.

[0121] Also within the scope of the present invention is apharmaceutical composition or formulation which comprises apharmaceutical carrier and at least one compound of formula I or itspharmaceutically acceptable salts or stereoisomers thereof.

[0122] The present invention further includes a process of making apharmaceutical composition or formulation comprising a compound offormula I or its pharmaceutically acceptable salt or stereoisomersthereof and a pharmaceutically acceptable carrier.

[0123] The present invention further includes a process of preparing acompound of formula I.

DETAILED DESCRIPTION OF THE INVENTION

[0124] The present invention relates to melanocortin receptor agonists,and more particularly piperazine derivatives as melanocortin receptoragonists. The compounds of present invention are useful for thetreatment or prevention of diseases and disorders responsive to theactivation of melanocortin receptors, such as obesity, diabetes andsexual dysfunction including erectile dysfunction and female sexualdysfunction.

[0125] An embodiment of the present invention is a compound of formulaI:

[0126] or a pharmaceutically acceptable salts or stereoisomers thereof,wherein

[0127] L and L¹ are independently: hydrogen or together oxo;

[0128] T is:

[0129] R is independently:

[0130] hydrogen,

[0131] hydroxy,

[0132] cyano,

[0133] nitro,

[0134] halo,

[0135] C₁-C₈ alkyl,

[0136] C₁-C₈ alkoxy,

[0137] C₁-C₄ haloalkyl,

[0138] (D)C(O)R⁹,

[0139] (D)C(O)OR⁹,

[0140] (D)C(O)SR⁹,

[0141] (D)C(O)heteroaryl,

[0142] (D)C(O)heterocyclyl,

[0143] (D)C(O)N(R⁹)₂,

[0144] (D))N(R⁹)₂,

[0145] (D)NR⁹COR⁹,

[0146] (D)NR⁹CON(R⁹)₂,

[0147] (D)NR⁹C(O)OR⁹,

[0148] (D)NR⁹C(R⁹)═N(R⁹),

[0149] (D)NR⁹C(═NR⁹)N(R⁹)₂,

[0150] (D)NR⁹SO₂R⁹,

[0151] (D)NR⁹SO₂N(R⁹)₂,

[0152] (D)NR⁹(CH₂)_(n)heterocyclyl,

[0153] (D)NR⁹(CH₂)_(n)heteroaryl,

[0154] (D)OR⁹,

[0155] OSO₂R⁹,

[0156] (D)[O]_(q)(C₃-C₇ cycloalkyl),

[0157] (D)[O]_(q)(CH₂)_(n)aryl,

[0158] (D)[O]_(q)(CH₂)_(n)heteroaryl,

[0159] (D)[O]_(q)(CH₂)_(n) heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen when q=1,

[0160] (D)SR⁹,

[0161] (D)SOR⁹,

[0162] (D)SO₂R⁹, or

[0163] (D)SO₂N(R⁹)₂;

[0164] wherein C₁-C₈ alkyl, C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl,heterocyclyl and heteroaryl are optionally substituted with one to fivesubstituents independently selected from R⁸;

[0165] R¹ is independently:

[0166] hydrogen, CONH(C₁-C₈ alkyl), C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇cycloalkyl or oxo, provided that oxo is not attached to the same carbonthat attached to nitrogen which forms an amide bond;

[0167] R³ is independently: aryl or thienyl;

[0168] wherein aryl and thienyl are optionally substituted with one tothree substituents selected from the group consisting of:

[0169] cyano, halo, C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy,C₁-C₄ haloalkyl and C₁-C₄ haloalkyloxy;

[0170] R⁴ is independently:

[0171] hydrogen, C₁-C₈ alkyl, C(O)R⁹, C(O)OR⁹, C₃-C₇ cycloalkyl or(CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;

[0172] each R⁸ is independently:

[0173] hydrogen,

[0174] halo,

[0175] oxo

[0176] N(R¹⁰)₂

[0177] C₁-C₈ alkyl,

[0178] (D)C₃-C₇ cycloalkyl,

[0179] C₁-C₄ haloalkyl,

[0180] C₁-C₄ alkoxy,

[0181] heteroaryl,

[0182] hydroxy,

[0183] heterocyclyl, wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen,

[0184] phenyl,

[0185] (D)COR⁹,

[0186] (D)C(O)OR⁹

[0187] (D)OR⁹,

[0188] (D)OCOR⁹,

[0189] (D)OCO₂R⁹,

[0190] (D)SR⁹,

[0191] (D)SOR⁹, or

[0192] (D)SO₂R⁹;

[0193] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0194] each R⁹ is independently:

[0195] hydrogen,

[0196] C₁-C₈ alkyl,

[0197] C₁-C₄ haloalkyl,

[0198] (D)C₃-C₇ cycloalkyl,

[0199] (D)aryl, wherein aryl being phenyl or naphthyl,

[0200] (D)heteroaryl or

[0201] (D)heterocyclyl; wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen; and

[0202] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0203] each R¹⁰ is independently:

[0204] hydrogen, (C₁-C₈)alkyl, C(O)C₁-C₈ alkyl, aryl or C₃-C₇cycloalkyl;

[0205] each R¹¹ is independently:

[0206] hydrogen,

[0207] C₁-C₈ alkyl,

[0208] (D)aryl,

[0209] (D)heteroaryl

[0210] (CH₂)_(n)N(R⁸)₂,

[0211] (CH₂)_(n)NR⁸C(O)C₁-C₄ alkyl,

[0212] (CH₂)_(n)NR⁸SO₂C₁-C₄ alkyl,

[0213] (CH₂)_(n)SO₂N(R⁸)₂,

[0214] (CH₂)_(n)[O]_(q)C₁-C₈ alkyl,

[0215] (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸COR⁸,

[0216] (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸SO₂R⁸,

[0217] (CH₂)_(n)[O]_(q)-heterocyclyl or

[0218] (CH₂)_(n)[O]_(q)(C₁-C₈ alkyl)-heterocyclyl; and

[0219] wherein n is 2-8;

[0220] each R¹² is independently:

[0221] hydrogen,

[0222] C₁-C₈ alkyl,

[0223] (D)phenyl

[0224] C(O)C₁-C₈ alkyl,

[0225] C(O)phenyl,

[0226] SO₂C₁-C₈ alkyl or

[0227] SO₂-phenyl;

[0228] D is a bond or —(CH₂)_(n)—;

[0229] n is 0-8;

[0230] p is 0-5;

[0231] q is 0-1; and

[0232] r is 1-2.

[0233] The compound of the present invention as recited above, whereinR³ is phenyl optionally para-substituted with chloro, bromo, fluoro,iodo, methoxy, benzyloxy or methyl. The preferred R³ is phenylpara-substituted with chloro, fluoro or methoxy.

[0234] The compound of the present invention as recited above, whereinR⁴ is hydrogen.

[0235] The compound of the present invention as recited above, wherein—(CH₂)_(n)—T is:

[0236] where * denotes a chiral carbon atom having a R or Sconfiguration.

[0237] The compound of the present invention as recited above, wherein Land L¹ are together oxo and the chiral carbon has R configuration.

[0238] The preferred embodiment of the present invention provides acompound of formula II,

[0239] or a pharmaceutically acceptable salts or stereoisomers thereof.

[0240] Yet another preferred embodiment of the present inventionprovides a compound of formula III,

[0241] or a pharmaceutically acceptable salts or stereoisomers thereof.

[0242] Yet another preferred embodiment of the present inventionprovides a compound of formula IV,

[0243] or a pharmaceutically acceptable salts or stereoisomers thereof.

[0244] Yet another preferred embodiment of the present inventionprovides a compound of formula V,

[0245] or a pharmaceutically acceptable salts or stereoisomers thereof.

[0246] The compound of the present invention as recited above in formulaII to V wherein,

[0247] P is 0-5;

[0248] n is 0-8;

[0249] q is 0-1;

[0250] D is a bond or —(CH₂)_(n)—;

[0251] R is independently:

[0252] hydrogen,

[0253] hydroxy,

[0254] cyano,

[0255] nitro,

[0256] halo,

[0257] C₁-C₈ alkyl,

[0258] C₁-C₈ alkoxy,

[0259] C₁-C₄ haloalkyl,

[0260] (D)C(O)R⁹,

[0261] (D)C(O)OR⁹,

[0262] (D)C(O)SR⁹,

[0263] (D)C(O)heteroaryl,

[0264] (D)C(O)heterocyclyl,

[0265] (D)C(O)N(R⁹)₂,

[0266] (D)N(R⁹)₂,

[0267] (D)NR⁹COR⁹,

[0268] (D)NR⁹CON(R⁹)₂,

[0269] (D)NR⁹C(O)OR⁹,

[0270] (D)NR⁹C(R⁹)═N(R⁹),

[0271] (D)NR⁹C(═NR⁹)N(R⁹)₂,

[0272] (D)NR⁹SO₂R⁹,

[0273] (D)NR⁹SO₂N(R⁹)₂,

[0274] (D)NR⁹(CH₂)_(n)heterocyclyl,

[0275] (D)NR⁹(CH₂)_(n)heteroaryl,

[0276] (D)OR⁹,

[0277] OSO₂R⁹,

[0278] (D)[O]_(q)(C₃-C₇ cycloalkyl),

[0279] (D)[O]_(q)(CH₂)_(n)aryl,

[0280] (D)[O]_(q)(CH₂)_(n)heteroaryl,

[0281] (D)[O]_(q)(CH₂)_(n) heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen when q=1,

[0282] (D)SR⁹,

[0283] (D)SOR⁹,

[0284] (D)SO₂R⁹, or

[0285] (D)SO₂N(R⁹)₂;

[0286] wherein C₁-C₈ alkyl, C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl,heterocyclyl and heteroaryl are optionally substituted with one to fivesubstituents independently selected from R⁸;

[0287] each R⁸ is independently:

[0288] hydrogen,

[0289] halo,

[0290] oxo

[0291] N(R¹⁰)₂

[0292] C₁-C₈ alkyl,

[0293] (D)C₃-C₇ cycloalkyl,

[0294] C₁-C₄ haloalkyl,

[0295] C₁-C₄ alkoxy,

[0296] heteroaryl,

[0297] hydroxy,

[0298] heterocyclyl, wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen,

[0299] phenyl,

[0300] (D)COR⁹,

[0301] (D)C(O)OR⁹

[0302] (D)OR⁹,

[0303] (D)OCOR⁹,

[0304] (D)OCO₂R⁹,

[0305] (D)SR⁹,

[0306] (D)SOR⁹, or

[0307] (D)SO₂R⁹;

[0308] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0309] each R⁹ is independently:

[0310] hydrogen,

[0311] C₁-C₈ alkyl,

[0312] C₁-C₄ haloalkyl,

[0313] (D)C₃-C₇ cycloalkyl,

[0314] (D)aryl, wherein aryl being phenyl or naphthyl heteroaryl or

[0315] heterocyclyl; wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen; and

[0316] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;and

[0317] each R¹⁰ is independently:

[0318] hydrogen, (C₁-C₈)alkyl, C(O)C₁-C₈ alkyl, aryl or C₃-C₇cycloalkyl.

[0319] The compound of the present invention as recited above in formulaIV, wherein R¹⁰ is hydrogen or (C₁-C₈)alkyl.

[0320] The most preferred compound of the present invention is thecompound listed below: Name of Compounds Structure of Compounds1-(D-Tic-4-Cl-D-Phe)-4-(2- methanesulfonylamino-phenyl)- piperazine

1-(D-Tic-4-Cl-D-Phe)-4-(2- dimethylaminomethyl-phenyl)- piperazine

3-(4-chloro-phenyl)-2-[(1,2,3,4- tetrahydro-isoquinolin-3-ylmethyl)-amino]-1-[4-(2-[1,2,4]triazol-1- ylmethyl-phenyl)-piperazin-1-yl]-propan-1-one

N-(1-(4-chloro-benzyl)-2-{4-[2- (isobutyl-methanesulfonyl-amino)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-2- (2,3-dihydro-1H-isoindol-1-yl)-acetamide

[0321] Also encompassed by the present invention is a pharmaceuticalcomposition or formulation, which comprises a pharmaceutical carrier andat least one compound of formula I or its pharmaceutically acceptablesalts or stereoisomers thereof. The pharmaceutical composition and orformulation may optionally further include a second active ingredientselected from the group consisting of an insulin sensitizer, insulinmimetic, sulfonylurea, alpha-glucosidase inhibitor, HMG-CoA reductaseinhibitor, sequestrant cholesterol lowering agent, beta 3 adrenergicreceptor agonist, neuropeptide Y antagonist, phosphodiester V inhibitor,and an alpha 2 adrenergic receptor antagonist.

[0322] Yet another aspect of the present invention is a process ofmaking a pharmaceutical composition comprising a compound of formula Ior its pharmaceutically acceptable salt or stereoisomers thereof asrecited above and a pharmaceutically acceptable carrier.

[0323] Yet another aspect of the present invention is a method ofpreventing or treating obesity or diabetes mellitus in mammal comprisingthe administration of a therapeutically effective amount of the compoundof formula I.

[0324] Yet anther aspect of the present invention is a method ofpreventing or treating male or female sexual dysfunction in mammal, morespecifically the male or female sexual dysfunction, comprising theadministration of a therapeutically effective amount of the compound offormula I.

[0325] Yet anther aspect of the present invention is a process forpreparing a compound of formula I,

[0326] or a pharmaceutically acceptable salts or stereoisomers thereof,wherein

[0327] —CLL¹—(CH₂)_(n)—T is:

[0328] wherein R₁ is hydrogen, C₁-C₈ alkyl, Boc, CBZ, phenyl, FMOC or(C₁-C₈ alkyl)phenyl;

[0329] Q represents a moiety:

[0330] R is independently:

[0331] hydrogen,

[0332] hydroxy,

[0333] cyano,

[0334] nitro,

[0335] halo,

[0336] C₁-C₈ alkyl,

[0337] C₁-C₈ alkoxy,

[0338] C₁-C₄ haloalkyl,

[0339] (D)C(O)R⁹,

[0340] (D)C(O)OR⁹,

[0341] (D)C(O)SR⁹,

[0342] (D)C(O)heteroaryl,

[0343] (D)C(O)heterocyclyl,

[0344] (D)C(O)N(R⁹)₂,

[0345] (D)N(R⁹)₂,

[0346] (D)NR⁹COR⁹,

[0347] (D)NR⁹CON(R⁹)₂,

[0348] (D)NR⁹C(O)OR⁹,

[0349] (D)NR⁹C(R⁹)═N(R⁹),

[0350] (D)NR⁹C(═NR⁹)N(R⁹)₂,

[0351] (D)NR⁹SO₂R⁹,

[0352] (D)NR⁹SO₂N(R⁹)₂,

[0353] (D)NR⁹(CH₂)_(n)heterocyclyl,

[0354] (D)NR⁹(CH₂)_(n)heteroaryl,

[0355] (D)OR⁹,

[0356] OSO₂R⁹,

[0357] (D)[O]_(q)(C₃-C₇ cycloalkyl),

[0358] (D)[O]_(q)(CH₂)_(n)aryl,

[0359] (D)[O]_(q)(CH₂)_(n)heteroaryl,

[0360] (D)[O]_(q)(CH₂)_(n) heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen when q=1,

[0361] (D)SR⁹,

[0362] (D)SOR⁹,

[0363] (D)SO₂R⁹, or

[0364] (D)SO₂N(R⁹)₂;

[0365] wherein C₁-C₈ alkyl, C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl,heterocyclyl and heteroaryl are optionally substituted with one to fivesubstituents independently selected from R⁸;

[0366] R¹ is independently:

[0367] hydrogen, CONH(C₁-C₈ alkyl), C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇cycloalkyl or oxo, provided that oxo is not attached to the same carbonthat attached to nitrogen which forms an amide bond;

[0368] R³ is independently: aryl or thienyl;

[0369] wherein aryl and thienyl are optionally substituted with one tothree substituents selected from the group consisting of:

[0370] cyano, halo, C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy,C₁-C₄ haloalkyl and C₁-C₄ haloalkyloxy;

[0371] R⁴ is independently:

[0372] hydrogen, C₁-C₈ alkyl, C(O)R⁹, C(O)OR⁹, C₃-C₇ cycloalkyl or(CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;

[0373] each R⁸ is independently:

[0374] hydrogen,

[0375] halo,

[0376] oxo

[0377] N(R¹⁰)₂

[0378] C₁-C₈ alkyl,

[0379] (D)C₃-C₇ cycloalkyl,

[0380] C₁-C₄ haloalkyl,

[0381] C₁-C₄ alkoxy,

[0382] heteroaryl,

[0383] hydroxy,

[0384] heterocyclyl, wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen,

[0385] phenyl,

[0386] (D)COR⁹,

[0387] (D)C(O)OR⁹

[0388] (D)OR⁹,

[0389] (D)OCOR⁹,

[0390] (D)OCO₂R⁹,

[0391] (D)SR⁹,

[0392] (D)SOR⁹, or

[0393] (D)SO₂R⁹;

[0394] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0395] each R⁹ is independently:

[0396] hydrogen,

[0397] C₁-C₈ alkyl,

[0398] C₁-C₄ haloalkyl,

[0399] (D)C₃-C₇ cycloalkyl,

[0400] (D)aryl, wherein aryl being phenyl or naphthyl,

[0401] (D)heteroaryl or

[0402] (D)heterocyclyl; wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen; and

[0403] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0404] each R¹⁰ is independently:

[0405] hydrogen, (C₁-C₈)alkyl, C(O)C₁-C₈ alkyl, aryl or C₃-C₇cycloalkyl;

[0406] D is a bond or —(CH₂)_(n)—;

[0407] n is 0-8;

[0408] p is 0-5;

[0409] q is 0-1; and

[0410] r is 1-2;

[0411] comprising the steps of:

[0412] a) reacting a compound having a structural formula 1,

[0413]  with CH₂CH═C(O)OR^(a) wherein R^(a) is hydrogen or C₁-C₈ alkyland X is halo, in the presence of a catalyst and a base in a suitableorganic solvent to give the compound of formula 2,

[0414] b) reductively aminating the compound of formula 2 in thepresence of amine in an acidic condition to give a compound of formula3,

[0415] c) cyclizing the compound of formula 3 by Michael addition togive a compound of formula 4 or stereoisomers thereof,

[0416] d) coupling the compound of formula 4 or stereoisomers thereof,wherein R^(a) of compound 4 is H, with a compound of formula 5,

[0417]  wherein R^(a) of compound 5 is C₁-C₈ alkyl, to give a compoundof formula 6;

[0418] e) coupling the compound of formula 6, wherein R^(a) is H, with acompound having a structural,

[0419]  to afford the compound of formula 1.

[0420] The process of the present invention as recited above, wherein

[0421] in Step (a) is 2-boromobenzaldehydes.

[0422] The process of the present invention as recited above, whereinCH₂CH═C(O)OR in Step (a) is methylacrylate.

[0423] The process of the present invention as recited above, whereinthe catalyst in Step (a) is selected from the group consisting of:Pd(Ph₃P)₂Cl₂, Pd(Ph₃P)₄Cl₂, Pd(Ph₃P)₄, Pd(Ph₃P)₂Cl₂/CuI,Pd(OAc)₂/Ph₃P-Bu₄NBr, Pd(Ph₃P)₄Cl₂/H₂ and Pd(OAc)₂/P(O-tol)₃; andwherein the base in Step (a) is NR₃ wherein R is hydrogen or C₁-C₈alkyl.

[0424] The process of the present invention as recited above, whereinthe amine in Step (b) is selected from the group consisting of:benzylamine, alpha-methylbenzylamine and BocNH₂.

[0425] The process of the present invention as recited above, whereinthe Step (b) further comprises reducing of intermediate imine compoundin the presence of reducing agent, the reducing agent being selectedfrom the group consisting of: NaCNBH₃, Na(OAc)₃BH, NaBH₄/H+, and acombination of Et₃SiH and TFA in CH₃CN or CH₂Cl₂.

[0426] The process of the present invention as recited above, whereinthe stereoisomer of compound of formula 4 in Step (c) is a compound offormula 4a.

[0427] The process of the present invention as recited above, whereinthe compound of formula 4a is prepared by asymmetric hydrogenation of acompound having structural formula,

[0428] The process of the present invention as recited above, whereinthe Michael addition in Step (c) is carried out in a basic workupcondition.

[0429] The process of the present invention as recited above, whereinthe Step (e) further comprises deprotecting or protecting of thecompound of formula (4) at NR₁.

[0430] Yet another aspect of the present invention is a process forpreparing a compound of formula I,

[0431] or a pharmaceutically acceptable salts or stereoisomers thereof,wherein

[0432] —CLL¹—(CH₂)_(n)—T is:

[0433] Q represents a moiety:

[0434] R is independently:

[0435] hydrogen,

[0436] hydroxy,

[0437] cyano,

[0438] nitro,

[0439] halo,

[0440] C₁-C₈ alkyl,

[0441] C₁-C₈ alkoxy,

[0442] C₁-C₄ haloalkyl,

[0443] (D)C(O)R⁹,

[0444] (D)C(O)OR⁹,

[0445] (D)C(O)SR⁹,

[0446] (D)C(O)heteroaryl,

[0447] (D)C(O)heterocyclyl,

[0448] (D)C(O)N(R⁹)₂,

[0449] (D)N(R⁹)₂,

[0450] (D)NR⁹COR⁹,

[0451] (D)NR⁹CON(R⁹)₂,

[0452] (D)NR⁹C(O)OR⁹,

[0453] (D)NR⁹C(R⁹)═N(R⁹),

[0454] (D)NR⁹C(═NR⁹)N(R⁹)₂,

[0455] (D)NR⁹SO₂R⁹,

[0456] (D)NR⁹SO₂N(R⁹)₂,

[0457] (D)NR⁹(CH₂)_(n)heterocyclyl,

[0458] (D)NR⁹(CH₂)_(n)heteroaryl,

[0459] (D)OR⁹,

[0460] OSO₂R⁹,

[0461] (D)[O]_(q)(C₃-C₇ cycloalkyl),

[0462] (D)[O]_(q)(CH₂)_(n)aryl,

[0463] (D)[O]_(q)(CH₂)_(n)heteroaryl,

[0464] (D)[O]_(q)(CH₂)_(n) heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen when q=1,

[0465] (D)SR⁹,

[0466] (D)SOR⁹,

[0467] (D)SO₂R⁹, or

[0468] (D)SO₂N(R⁹)₂;

[0469] wherein C₁-C₈ alkyl, C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl,heterocyclyl and heteroaryl are optionally substituted with one to fivesubstituents independently selected from R⁸;

[0470] R¹ is independently:

[0471] hydrogen, CONH(C₁-C₈ alkyl), C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇cycloalkyl or oxo, provided that oxo is not attached to the same carbonthat attached to nitrogen which forms an amide bond;

[0472] R³ is independently: aryl or thienyl;

[0473] wherein aryl and thienyl are optionally substituted with one tothree substituents selected from the group consisting of:

[0474] cyano, halo, C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy,C₁-C₄ haloalkyl and C₁-C₄ haloalkyloxy;

[0475] R⁴ is independently:

[0476] hydrogen, C₁-C₈ alkyl, C(O)R⁹, C(O)OR⁹, C₃-C₇ cycloalkyl or(CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;

[0477] each R⁸ is independently:

[0478] hydrogen,

[0479] halo,

[0480] oxo

[0481] N(R¹⁰)₂

[0482] C₁-C₈ alkyl,

[0483] (D)C₃-C₇ cycloalkyl,

[0484] C₁-C₄ haloalkyl,

[0485] C₁-C₄ alkoxy,

[0486] heteroaryl,

[0487] hydroxy,

[0488] heterocyclyl, wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen,

[0489] phenyl,

[0490] (D)COR⁹,

[0491] (D)C(O)OR⁹

[0492] (D)OR⁹,

[0493] (D)OCOR⁹,

[0494] (D)OCO₂R⁹,

[0495] (D)SR⁹,

[0496] (D)SOR⁹, or

[0497] (D)SO₂R⁹;

[0498] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0499] each R⁹ is independently:

[0500] hydrogen,

[0501] C₁-C₈ alkyl,

[0502] C₁-C₄ haloalkyl,

[0503] (D)C₃-C₇ cycloalkyl,

[0504] (D)aryl, wherein aryl being phenyl or naphthyl,

[0505] (D)heteroaryl or

[0506] (D)heterocyclyl; wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen; and

[0507] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0508] each R¹⁰ is independently:

[0509] hydrogen, (C₁-C₈)alkyl, C(O)C₁-C₈ alkyl, aryl or C₃-C₇cycloalkyl;

[0510] each R¹¹ is independently: hydrogen or (C₁-C₈)alkyl;

[0511] D is a bond or —(CH₂)_(n)—;

[0512] n is 0-8;

[0513] p is 0-5;

[0514] q is 0-1; and

[0515] r is 1-2;

[0516] comprising the steps of:

[0517] a) esterifying a compound of formula 1,

[0518]  with an alcohol R^(a)OH to form a compound of formula 2,

[0519]  wherein R^(a) is C₁-C₄ alkyl or (D)phenyl;

[0520] b) reacting a compound of formula 2 with R¹¹COR¹¹ to form acompound of formula 3,

[0521]  wherein R¹¹ is independently hydrogen or C₁-C₄ alkyl;

[0522] c) reacting a compound of formula 3 with an activating group toform a compound of formula 4,

[0523]  wherein A is an activating group;

[0524] d) deoxygenating the compound of formula 4 by hydrogenation toafford a compound of formula 5,

[0525] e) optionally reacting the compound of formula 5 with aninorganic base to form a compound of formula 6,

[0526]  wherein HA is an acidic and M is a univalent cation;

[0527] f) resolving the compound of formula 5 or formula 6 to afford achiral compound of formula 7,

[0528]  wherein M is hydrogen and R^(a′) is H or R^(a);

[0529] g) coupling the compound of formula 7 with a compound of formula8,

[0530]  to afford a compound of formula 9,

[0531] h) coupling the compound of formula 9 with a compound having aformula,

[0532]  to afford a compound of formula I.

[0533] Yet another aspect of the present invention is a process forpreparing a compound of formula I,

[0534] or a pharmaceutically acceptable salts or stereoisomers thereof,wherein

[0535] —CLL¹—(CH₂)_(n)—T is:

[0536] Q represents a moiety:

[0537] R is independently:

[0538] hydrogen,

[0539] hydroxy,

[0540] cyano,

[0541] nitro,

[0542] halo,

[0543] C₁-C₈ alkyl,

[0544] C₁-C₈ alkoxy,

[0545] C₁-C₄ haloalkyl,

[0546] (D)C(O)R⁹,

[0547] (D)C(O)OR⁹,

[0548] (D)C(O)SR⁹,

[0549] (D)C(O)heteroaryl,

[0550] (D)C(O)heterocyclyl,

[0551] (D)C(O)N(R⁹)₂,

[0552] (D)N(R⁹)₂,

[0553] (D)NR⁹COR⁹,

[0554] (D)NR⁹CON(R⁹)₂,

[0555] (D)NR⁹C(O)OR⁹,

[0556] (D)NR⁹C(R⁹)═N(R⁹),

[0557] (D)NR⁹C(═NR⁹)N(R⁹)₂,

[0558] (D)NR⁹SO₂R⁹,

[0559] (D)NR⁹SO₂N(R⁹)₂,

[0560] (D)NR⁹(CH₂)_(n)heterocyclyl,

[0561] (D)NR⁹(CH₂)_(n)heteroaryl,

[0562] (D)OR⁹,

[0563] OSO₂R⁹,

[0564] (D)[O]_(q)(C₃-C₇ cycloalkyl),

[0565] (D)[O]_(q)(CH₂)_(n)aryl,

[0566] (D)[O]_(q)(CH₂)_(n)heteroaryl,

[0567] (D)[O]_(q)(CH₂)_(n) heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen when q=1,

[0568] (D)SR⁹,

[0569] (D)SOR⁹,

[0570] (D)SO₂R⁹, or

[0571] (D)SO₂N(R⁹)₂;

[0572] wherein C₁-C₈ alkyl, C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl,heterocyclyl and heteroaryl are optionally substituted with one to fivesubstituents independently selected from R⁸;

[0573] R¹ is independently:

[0574] hydrogen, CONH(C₁-C₈ alkyl), C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇cycloalkyl or oxo, provided that oxo is not attached to the same carbonthat attached to nitrogen which forms an amide bond;

[0575] R³ is independently: aryl or thienyl;

[0576] wherein aryl and thienyl are optionally substituted with one tothree substituents selected from the group consisting of:

[0577] cyano, halo, C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy,C₁-C₄ haloalkyl and C₁-C₄ haloalkyloxy;

[0578] R⁴ is independently:

[0579] hydrogen, C₁-C₈ alkyl, C(O)R⁹, C(O)OR⁹, C₃-C₇ cycloalkyl or(CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;

[0580] each R⁸ is independently:

[0581] hydrogen,

[0582] halo,

[0583] oxo

[0584] N(R¹⁰)₂

[0585] C₁-C₈ alkyl,

[0586] (D)C₃-C₇ cycloalkyl,

[0587] C₁-C₄ haloalkyl,

[0588] C₁-C₄ alkoxy,

[0589] heteroaryl,

[0590] hydroxy,

[0591] heterocyclyl, wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen,

[0592] phenyl,

[0593] (D)COR⁹,

[0594] (D)C(O)OR⁹

[0595] (D)OR⁹,

[0596] (D)OCOR⁹,

[0597] (D)OCO₂R⁹,

[0598] (D)SR⁹,

[0599] (D)SOR⁹, or

[0600] (D)SO₂R⁹;

[0601] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0602] each R⁹ is independently:

[0603] hydrogen,

[0604] C₁-C₈ alkyl,

[0605] C₁-C₄ haloalkyl,

[0606] (D)C₃-C₇ cycloalkyl,

[0607] (D)aryl, wherein aryl being phenyl or naphthyl,

[0608] (D)heteroaryl or

[0609] (D)heterocyclyl; wherein heterocyclyl excludes a heterocyclylcontaining a single nitrogen; and

[0610] wherein aryl, heteroaryl, heterocyclyl, alkyl or cycloalkyl isoptionally substituted with one to three substituents selected from thegroup consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰;

[0611] each R¹⁰ is independently:

[0612] hydrogen, (C₁-C₈)alkyl, C(O)C₁-C₈ alkyl, aryl or C₃-C₇cycloalkyl;

[0613] each R¹⁰ is independently: hydrogen or (C₁-C₈)alkyl;

[0614] D is a bond or —(CH₂)_(n)—;

[0615] n is 0-8;

[0616] p is 0-5;

[0617] q is 0-1; and

[0618] r is 1-2;

[0619] comprising the steps of:

[0620] a) reacting a compound formula 1:

[0621]  wherein X is halo, and R¹¹ is independently, hydrogen or C₁-C₄alkyl, with CNCH₂CO₂R^(a) wherein R^(a) is C₁-C₈ alkyl or benzyl toafford a compound of formula 2:

[0622] b) protecting the compound of formula 2 to form the compound offormula 3:

[0623] c) hydrogenating the compound of formula 3 to afford a compoundof formula 4:

[0624] d) coupling the compound of formula 4 wherein R^(a′) is hydrogenor R^(a), with a compound of formula 5,

[0625]  to afford a compound of formula 6,

[0626] e) coupling the compound of formula 6 with a compound having aformula,

[0627]  Q to afford a compound of formula I.

[0628] Throughout the instant application, the following terms have theindicated meanings:

[0629] The term “alkyl,” unless otherwise indicated, refers to thosealkyl groups of a designated number of carbon atoms of either a straightor branched saturated configuration. Examples of “alkyl” includes, butare not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, sec-butyl and t-butyl, pentyl, hexyl, neopenyl, isopentyl andthe like. Alkyl as defined above may be optionally substituted with adesignated number of substituents as set forth in the embodiment recitedabove.

[0630] The term “alkenyl” means hydrocarbon chain of a specified numberof carbon atoms of either a straight or branched configuration andhaving at least one carbon-carbon double bond, which may occur at anypoint along the chain, such as ethenyl, propenyl, butenyl, pentenyl,vinyl, alkyl, 2-butenyl and the like. Alkenyl as defined above may beoptionally substituted with designated number of substituents as setforth in the embodiment recited above.

[0631] The term “haloalkyl” is an alkyl group of indicated number ofcarbon atoms, which is substituted with one to five halo atoms selectedfrom F, Br, Cl and I. An example of a haloalkyl group istrifluoromethyl.

[0632] The term “alkoxy” represents an alkyl group of indicated numberof carbon atoms attached through an oxygen bridge, such as methoxy,ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentoxy, and the like.Alkoxy as defined above may be optionally substituted with a designatednumber of substituents as set forth in the embodiment recited above.

[0633] The term “cycloalkyl” refers to a ring composed of 3 to 7methylene groups, each of which may be optionally substituted with otherhydrocarbon substituents. Examples of cycloalkyl includes, but are notlimited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl andcycloheptyl, and the like. Cycloalkyl as defined above may be optionallysubstituted with a designated number of substituents as set forth in theembodiment recited above.

[0634] The term “halo” refers to fluoro, chloro, bromo and iodo.

[0635] The term “haloalkyloxy” represents a haloalkyl group of indicatednumber of carbon atoms attached through an oxygen bridge, such as OCF₃.“Haloalkyloxy” as defined above may be optionally substituted with adesignated number of substituents as set forth in the embodiment recitedabove.

[0636] The term “aryl” refers to phenyl, naphthyl, anthracenyl,phenanthrenyl and the like which is optionally substituted with adesignated number of substituents as set forth in the embodiment recitedabove.

[0637] The term “heteroaryl” refers to monocyclic or bicyclic aromaticring of 5- to 10-carbon atoms containing from one to four heteroatomsselected from O, N, or S, and the heteroaryl being optionallysubstituted with a designated number of substituents as set forth in theembodiment recited above. Examples of heteroaryl are, but are notlimited to furanyl, thienyl, thiazolyl, imidazolyl, isoxazoyl, oxazoyl,pyrazoyl, pyrrolyl, pyrazinyl, pyridyl, pyrimidyl, and purinyl,cinnolinyl, benzothienyl, benzotriazolyl, benzoxazolyl, quinoline,isoquinoline and the like.

[0638] The “heterocyclyl” is defined as a monocyclic, bicyclic, ortricyclic ring of 5 to 14 carbon atoms which are saturated or partiallysaturated containing from one to four heteroatoms selected from N, O orS. The “heterocycly” includes “nitrogen containing heterocyclyl,” whichcontains from one to four nitrogen atoms and optionally further containsone other heteroatom selected from O or S. Heterocyclyl as defined abovemay be optionally substituted with a designated number of substituentsas set forth in the embodiment recited above.

[0639] A mammal as used in here includes a human and a warm-bloodedanimal such as a cat, a dog and the like.

[0640] The term “composition” or “formulation”, as in pharmaceuticalcomposition or formulation, is intended to encompass a productcomprising the active ingredient(s), and the inert ingredient(s) thatmake up the carrier. Accordingly, the pharmaceutical compositions of thepresent invention encompass any composition made by admixing a compoundof the present invention (a compound of formula I) and apharmaceutically acceptable carrier.

[0641] The term “pharmaceutical” when used herein as an adjective meanssubstantially non-deleterious to the recipient mammal.

[0642] The term “unit dosage form” refers to physically discrete unitssuitable as unitary dosages for human subjects and other non-humananimals such as warm-blooded animals each unit containing apredetermined quantity of active ingredient (a compound of formula I)calculated to produce the desired therapeutic effect in association witha suitable pharmaceutical carrier.

[0643] The term “treating” or “preventing” as used herein includes itsgenerally accepted meanings, i.e., preventing, prohibiting, restraining,alleviating, ameliorating, slowing, stopping, or reversing theprogression or severity of a pathological condition, or sequela thereofas described herein.

[0644] “Erectile dysfunction” is a disorder involving the failure of amale mammal to achieve erection, ejaculation, or both. Symptoms oferectile dysfunction include an inability to achieve or maintain anerection, ejaculatory failure, premature ejaculation, inability toachieve an orgasm. An increase in erectile dysfunction is oftenassociated with age and is generally caused by a physical disease or asa side effect of drug treatment.

[0645] “Female sexual dysfunction” encompasses, without limitation,conditions such as a lack of sexual desire and related arousaldisorders, inhibited orgasm, lubrication difficulties, and vaginismus.

[0646] Because certain compounds of the invention contain an acidicmoiety (e.g., carboxy), the compound of formula I may exist as apharmaceutical base addition salt thereof. Such salts include thosederived from inorganic bases such as ammonium and alkali and alkalineearth metal hydroxides, carbonates, bicarbonates and the like, as wellas salts derived from basic organic amines such as aliphatic andaromatic amines, aliphatic diamines, hydroxy alkamines, and the like.

[0647] Because certain compounds of the invention contain a basic moiety(e.g., amino), the compound of formula I can also exist as apharmaceutical acid addition salt Such salts include sulfate,pyrosulfate, bisulfate, sulfite, bisulfite, phosphate,mono-hydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, chloride, bromide, iodide, acetate, propionate,decanoate, caprylate, acrylate, formate, isobutyrate, heptanoate,propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate,maleate, 2-butyne-1,4 dioate, 3-hexyne-2,5-dioate, benzoate,chlorobenzoate, hydroxybenzoate, methoxybenzoate, phthalate,xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate,citrate, lactate, hippurate, beta-hydroxybutyrate, glycollate, maleate,tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate,naphthalene-2-sulfonate, mandelate and the like salts. Preferred saltform of compound of formula I is an acid addition salts, morespecifically hydrochloride salt.

[0648] Some of the compounds described herein may exist as tautomerssuch as ketoenol tautomers. The individual tautomers as well as mixturesthereof are encompassed within the scope of the present invention.

[0649] Utility

[0650] Compounds of formula I are effective as melanocortin receptormodulators, particularly as agonists of the human MC-4 receptor. Asmelanocortin receptor agonists, the compounds of formula I are useful inthe treatment of diseases, disorders or conditions responsive to theactivation of one or more of the melanocortin receptors including, butnot limited to, MC-1, MC-2, MC-3, MC-4, and MC-5. Diseases, disorders orconditions receptive to treatment with a MC-4 agonist include thosementioned above and those described in WO 00/74679, the teachings ofwhich are herein incorporated by reference. In particular diseases,disorders or conditions receptive to treatment with a MC-4 agonistinclude obesity or diabetes mellitus, male or female sexual dysfunction,more specifically erectile dysfunction.

[0651] When describing various aspects of the present compounds offormula I, the terms “A domain”, “B domain” and “C domain” are usedbelow. This domain concept is illustrated below:

[0652] The following listing provides some of examples “A domain”, “Bdomain” and “C domain” of the compound of formula I. These listings areprovided as illustrative purposes and as such are not meant to belimiting.

[0653] Examples of A Domain:

[0654] Examples of B Domain:

[0655] Examples of C Domain:

[0656] Formulation

[0657] The compound of formula I is preferably formulated in a unitdosage form prior to administration. Accordingly the present inventionalso includes a pharmaceutical composition comprising a compound offormula I and a suitable pharmaceutical carrier.

[0658] The present pharmaceutical compositions are prepared by knownprocedures using well-known and readily available ingredients. In makingthe formulations of the present invention, the active ingredient (acompound of formula I) is usually be mixed with a carrier, or diluted bya carrier, or enclosed within a carrier, which may be in the form of acapsule, sachet, paper or other container. When the carrier serves as adiluent, it may be a solid, semisolid or liquid material which acts as avehicle, excipient or medium for the active ingredient. Thus, thecompositions can be in the form of tablets, pills, powders, lozenges,sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups,aerosol (as a solid or in a liquid medium), soft and hard gelatincapsules, suppositories, sterile injectable solutions and sterilepackaged powders.

[0659] Some examples of suitable carriers, excipients, and diluentsinclude lactose, dextrose, sucrose, sorbitol, mannitol, starches, gumacacia, calcium phosphate, alginates, tragacanth, gelatin, calciumsilicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose,water syrup, methyl cellulose, methyl and propylhydroxybenzoates, talc,magnesium stearate and mineral oil. The formulations can additionallyinclude lubricating agents, wetting agents, emulsifying and suspendingagents, preserving agents, sweetening agents or flavoring agents. Thecompositions of the invention may be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient.

[0660] Dosage:

[0661] The specific dose administered is determined by the particularcircumstances surrounding each situation. These circumstances include,the route of administration, the prior medical history of the recipient,the pathological condition or symptom being treated, the severity of thecondition/symptom being treated, and the age and sex of the recipient.Additionally, it would be understood that the therapeutic dosageadministered can be determined by the physician in the light of therelevant circumstances.

[0662] Generally, an effective minimum daily dose of a compound offormula I is about 1, 5, 10, 15, or 20 mg. Typically, an effectivemaximum dose is about 500, 100, 60, 50, or 40 mg. The suitable dose maybe determined in accordance with the standard practice in the medicalarts of “dose titrating” the recipient, which involves administering alow dose of the compound initially and then gradually increasing thedoes until the desired therapeutic effect is observed.

[0663] Route of Administration

[0664] The compounds may be administered by a variety of routesincluding the oral, rectal, transdermal, subcutaneous, topical,intravenous, intramuscular or intranasal routes.

[0665] Combination Therapy

[0666] Compounds of formula I may be used in combination with otherdrugs that are used in the treatment of the diseases or conditions forwhich compounds of formula I are useful. Such other drugs may beadministered by a route and in an amount commonly used therefor,contemporaneously or sequentially with a compound of formula I. When acompound of formula I is used contemporaneously with one or more otherdrugs, a pharmaceutical composition containing such other drugs inaddition to the compound of formula I is preferred. Accordingly, thepharmaceutical compositions of the present invention include those thatalso contain one or more other active ingredients in addition to acompound of formula I. Examples of other active ingredients that may becombined with a compound of formula I, either administered separately orin the same pharmaceutical compositions, include but are not limited to:

[0667] (a) insulin sensitizers including (i) PPARγ agonists such as theglitazones (e.g., troglitazone, pioglitazone, englitazone, MCC-555,BRL49653 and the like) and compounds disclosed in WO97/27857, WO97/28115, WO 97/28137 and WO97/27847; (ii) biguanides such as metforminand phenformin;

[0668] (b) insulin or insulin mimetics;

[0669] (c) sulfonylureas such as tolbutamide and glipizide;

[0670] (d) α-glucosidase inhibitors (such as acarbose),

[0671] (e) cholesterol lowering agents such as (i) HMG-CoA reductaseinhibitors (lovastatin, simvastatin and pravastatin, fluvastatin,atorvastatin, and other statins), (ii) sequestrants (cholestyramine,colestipol and a dialkylaminoalkyl derivatives of a cross-linkeddextran), (iii) nicotinyl alcohol nicotinic acid or a salt thereof, (iv)proliferator-activater receptor α-agonists such as fenofibric acidderivatives (gemfibrozil, clofibrat, fenofibrate and benzafibrate), (v)inhibitors of cholesterol absorption such as β-sitosterol and acylCoA:cholesterol acyltransferase inhibitors such as melinamide, (vi)probucol, (vii) vitamin E, and (viii) thyrornimetics;

[0672] (f) PPARδ agonists such as those disclosed in WO97/28149;

[0673] (g) antiobesity compounds such as fenfluramine, dexfenfluramine,phentermine, sibutramine, orlistat, and β-3 adrenergic receptoragonists;

[0674] (h) feeding behavior modifying agents such as neuropeptide Yantagonists (e.g., neuropeptide Y5) as disclosed in WO 97/19682, WO97/20820, WO 97/20821, WO 97/20822 and WO 97/20823;

[0675] (i) PPARα agonists as described in WO 97/36579;

[0676] (j) PPARγ antagonists as described in WO97/10813;

[0677] (k) serotonin reuptake inhibitors, such as fluoxetine andsertraline;

[0678] (l) growth hormone secretagogues such as MK-0677; and

[0679] (m) agents useful in the treatment of male and/or female sexualdysfunction, such as phosphodiester V inhibitors including sildenafiland ICI-351, and α-2 adrenergic receptor antagonists includingphentolamine mesylate; and dopamine-receptor agonists, such asapomorphine.

[0680] Biological Assays

[0681] A. Binding Assay:

[0682] The radioligand binding assay is used to identify competitiveinhibitors of ¹²⁵I-NDP-α-MSH binding to cloned human MCRs usingmembranes from stably transfected human embryonic kidney (HEK) 293cells.

[0683] HEK 293 cells transfected with human or rat melanocortininreceptors are grown either as adherent monolayers or suspension culture.Monolayer cells are grown in roller bottle cultures at 37° C. and 5%CO₂/air atmosphere in a 3:1 mixture of Dulbecco's modified Eagle medium(DMEM) and Ham's F12 containing 25 mM L-glucose, 100 units/ml penicillinG, 100 microgram/ml streptomyocin, 250 nanogram/ml amphoterin B, 300microgram/ml genticin and supplemented with 5% fetal bovine serum.Monolayer cells are adapted to suspension culture (Berg et al.,Biotechniques Vol. 14, No. 6, 1993) and are grown in either spinner orshaker flasks (37° C. and 7.5% CO₂/air overlay) in a modified DME/F12medium containing 0.1 mM CaCl₂, 2% equine serum and 100 microgram/mlsodium heparin to prevent cell-cell aggregation. Cells are harvested bycentrifugation, washed in PBS, and pellets are stored frozen at −80° C.until membrane preparations.

[0684] The cell pellets are resuspended in 10 volumes of membranepreparation buffer (i.e., 1 g pellet to 10 ml buffer) having thefollowing composition: 50 mM Tris pH 7.5 @ 4° C., 250 mM sucrose, 1 mMMgCl₂, Complete® EDTA-free protease inhibitor tablet (BoehringerMannheim), and 24 micrograms/ml DNase I (Sigma, St. Louis, Mo.). Thecells are homogenized with a motor-driven dounce using 20 strokes, andthe homogenate is centrifuged at 38,000×g at 4° C. for 40 minutes. Thepellets are resuspended in membrane preparation buffer at aconcentration of 2.5-7.5 mg/ml and 1 milliliter aliquots of membranehomogenates are quickly frozen in liquid nitrogen and then stored at−80° C.

[0685] Solutions of a compound of formula I (300 picomolar to 30micromolar) or unlabelled NDP-α-MSH (1 picomolar to 100 nanomolar) areadded to 150 microliters of membrane binding buffer to yield finalconcentrations (listed in parantheses). The membrane binding buffer hasthe following composition: 25 mM HEPES pH 7.5; 10 mM CaCl₂; 0.3% BSA).One hundred fifty microliters of membrane binding buffer containing0.5-5.0 microgram membrane protein is added, followed by 50 nanomolar¹²⁵I-NDP-α-MSH to final concentration of 100 picomolar. Additionally,fifty microliters of SPA beads (5 mg/ml) are added and the resultingmixture is agitated briefly and incubated for 10 hours at r.t. Theradioactivity is quantified in a Wallac Trilux Microplate Scintillationcounter. IC₅₀ values obtained in competition assays are converted toaffinity constants (K_(i) values) using the Cheng-Prusoff equation:K_(i)=IC₅₀/(1+D/K_(d)).

[0686] B. Functional Assay:

[0687] Functional cell based assays are developed to discriminateagonists and antagonists.

[0688] Agonist Assay: HEK 293 cells stably expressing a humanmelanocortin receptor (see e.g., Yang, et al., Mol-Endocrinol., 11(3):274-80, 1997) are dissociated from tissue culture flasks using atrypsin/EDTA solution(0.25%; Life Technologies, Rockville, Md.). Cellsare collected by centrifugation and resuspended in DMEM (LifeTechnologies, Rockville, Md.) supplemented with 1% L-glutamine and 0.5%fetal bovine serum. Cells are counted and diluted to 4.5×10⁵/ml.

[0689] A compound of formula I is diluted in dimethylsulfoxide (DMSO)(3×10⁻⁵ to 3×10³¹ ¹⁰ M final concentrations) and 0.05 volume of compoundsolution is added to 0.95 volumes of cell suspension; the final DMSOconcentration is 0.5%. After incubation at 37° C./5% CO₂ for 5 hours,cells are lysed by addition of luciferin solution (50 mM Tris, 1 mMMgCl₂, 0.2% Triton-X100, 5 mM DTT, 500 micromolar Coenzyme A, 150micromolar ATP, and 440 micromolar luciferin) to quantify the activityof the reporter gene luciferase, an indirect measurement ofintracellular cAMP production.

[0690] Luciferase activity is measured from the cell lysate using aWallac Victor 2 luminometer. The amount of lumen production whichresults from a compound of formula I is compared to that amount oflumens produced in response to NDP-α-MSH, defined as a 100% agonist, toobtain the relative efficacy of a compound. The EC₅₀ is defined as thecompound concentration that results in half maximal stimulation, whencompared to its own maximal level of stimulation.

[0691] Antagonist assay: Antagonist activity is defined as the abilityof a compound to block lumen production in response to NDP-α-MSH.Concentration-response curves are generated for NDP-α-MSH in the absenceand presence of a fixed concentration of a solution of a compound offormula I (10×K_(i) from binding assays). Suspensions of MCR-expressingcells are prepared and are incubated with NDP-α-MSH and compoundsolutions for 5 hours as described above. The assay is terminated by theaddition of luciferin reagent and lumen production is quantified.Antagonist potency is determined from the rightward shift of the EC₅₀value in the absence of a compound of formula I using the equation:K_(b)=Concentration of Antagonist/[(EC₅₀′/EC₅₀)−1].

[0692] Whole Cell cAMP Accumulation Assay

[0693] Compound Preparation

[0694] In the agonist assay, the compounds are prepared as 10 mM andNDP-alpha-MSH (control) as 33.3 μM stock solutions in 100% DMSO. Theseare serially diluted in 100% DMSO. The compound plate is further diluted1:200 in compound dilution buffer (HBSS-092, 1 mM Ascorbic Acid, 1 mMIBMX, 0.6% DMSO, 0.1% BSA). The final concentration range being 10μM-100 pM for compound and 33.33 nM-0.3 pM for control in 0.5% DMSO.Transfer 20 μl from this plate into four PET 96-well plates (all assaysare performed in duplicate for each receptor).

[0695] Cell Culture and Cell Stimulation

[0696] HEK 293 cells stably transfected with the MC3R and MC4R weregrown in DMEM containing 10% FBS and 1% Antibiotic/Antimycotic Solution.On the day of the assay the cells were dislodged with enzyme free celldissociation solution and resuspended in cell buffer (HBSS-092, 0.1%BSA, 10 mM HEPES) at 1×e6 cells/ml. Add 40 μl of cells/well to the PET96-well plates containing 20 microliter diluted compound and control.Incubate @ 37° C. in a waterbath for 20 minutes. Stop the assay byadding 50 μl Quench Buffer (50 mM Na Acetate, 0.25% Triton X-100).

[0697] Radioligand Binding Assays

[0698] Radioligand binding assays were run in SPA buffer (50 mM SodiumAcetate, 0.1% BSA). The beads, antibody and radioligand were diluted inSPA buffer to provide sufficient volume for each 96-well plate. To eachquenched assay well was added 100 microliter cocktail containing 33.33microliter of beads, 33.33 microliter antibody and 33.33 microliter¹²⁵I-cAMP. This was based on a final concentration of 6.3 mg/ml beads,0.65% anti-goat antibody and 61 pM of ¹²⁵I-cAMP (containing 25000-30000CPM) in a final assay volume of 210 microliter. The plates were countedin a Wallac MicroBeta counter after a 12-hour incubation.

[0699] The data was converted to pmoles cAMP using a standard curveassayed under the same conditions. The data was analyzed using ActivityBase software to generate agonist potencies (EC50) and percent relativeefficacy data to NDP-alpha-MSH.

[0700] C. In Vivo Food Intake Models:

[0701] 1) Daily food intake. Male Long-Evans rats are injectedintracerebroventricularly (ICV) with a test compound in 5 microliters of50% propylene glyco/artificial cerebrospinal fluid one hour prior toonset of dark cycle (12 hours). Food intake is determined by subtractingthe food weight remaining after 24 hours from food weight just prior toICV injection.

[0702] 2) Acute Calorimetry. Male Long-Evans rats are administered testcompound by subcutaneous injection, intramuscular injection, intravenousinjection, intraperitoneal injection, ICV injection or by oral gavagebetween 0 and 5 hours after the onset of the dark cycle. Rats are placedinto a calorimetry chamber and the volume of oxygen consumed and volumeor carbon dioxide exhaled are measured each hour for 24 hours. Foodintake is measured for the 24 hour period as described in C.1).Locomoter activity is measured when the rat breaks a series of infraredlaser beams when in the calorimeter. These measurements permitcalculation of energy expenditure, respiratory quotient and energybalance.

[0703] 3) Food intake in diet induced obese mice. Male C57/B16J micemaintained on a high fat diet (60% fat calories) for 6.5 months from 4weeks of age are dosed intraperitoneally with a compound of formula I.Food intake and body weight are measured over an eight day period.Biochemical parameters relating to obesity, including leptin, insulin,triglyceride, free fatty acid, cholesterol and serum glucose levels aredetermined.

[0704] D. Rat Ex Copula Assay:

[0705] Sexually mature male Caesarian Derived Sprague Dawley (CD) rats(over 60 days old) are used with the suspensory ligament surgicallyremoved to prevent retraction of the penis back into the penile sheathduring the ex copula evaluations. Animals receive food and water ad liband are kept on a normal light/dark cycle. Studies are conducted duringthe light cycle.

[0706] 1) Conditioning to Supine Restraint for Ex Copula Reflex Tests.This conditioning takes about 4 days. Day 1, the animals are placed in adarkened restrainer and left for 15-30 minutes. Day 2, the animals arerestrained in a supine position in the restrainer for 15-30 minutes. Day3, the animals are restrained in the supine position with the penilesheath retracted for 15-30 minutes. Day 4, the animals are restrained inthe supine position with the penile sheath retracted until penileresponses are observed. Some animals require additional days ofconditioning before they are completely acclimated to the procedures;non-responders are removed from further evaluation. After any handlingor evaluation, animals are given a treat to ensure positivereinforcement.

[0707] 2) Ex Copula Reflex Tests. Rats are gently restrained in a supineposition with their anterior torso placed inside a cylinder of adequatesize to allow for normal head and paw grooming. For a 400-500 gram rat,the diameter of the cylinder is approximately 8 cm. The lower torso andhind limbs are restrained with a non-adhesive material (vetrap). Anadditional piece of vetrap with a hole in it, through which the glanspenis will be passed, is fastened over the animal to maintain thepreputial sheath in a retracted position. Penile responses will beobserved, typically termed ex copulu genital reflex tests. Typically, aseries of penile erections will occur spontaneously within a few minutesafter sheath retraction. The types of normal reflexogenic erectileresponses include elongation, engorgement, cup and flip. An elongationis classified as an extension of the penile body. Engorgement is adilation of the glans penis. A cup is defined as an intense erectionwhere the distal margin of the glans penis momentarily flares open toform a cup. A flip is a dorsiflexion of the penile body.

[0708] Baseline and/or vehicle evaluations are conducted to determinehow and if an animal will respond. Some animals have a long durationuntil the first response while others are non-responders altogether.During this baseline evaluation, latency to first response time, numberand type of responses are recorded. The testing time frame is 15 minutesafter the first response.

[0709] After a minimum of 1 day between evaluations, these same animalsare administered a compound of formula I at 20 mg/kg and evaluated forpenile reflexes. All evaluations are videotaped and scored later. Dataare collected and analyzed using paired 2 tailed t-tests to comparedbaseline and/or vehicle evaluations to drug treated evaluations forindividual animals. Groups of a minimum of 4 animals are utilized toreduce variability.

[0710] Positive reference controls are included in each study to assurethe validity of the study. Animals can be dosed by a number of routes ofadministration depending on the nature of the study to be performed. Theroutes of administration includes intravenous (IV), intraperitoneal(IP), subcutaneous (SC) and intracerebral ventricular (ICY).

[0711] E. Models of Female Sexual Dysfunction:

[0712] Rodent assays relevant to female sexual receptivity include thebehavioral model of lordosis and direct observations of copulatoryactivity. There is also a urethrogenital reflex model in anesthetizedspinally transected rats for measuring orgasm in both male and femalerats. These and other established animal models of female sexualdysfunction are described in McKenna, et al., Am. J. Physiol.,(Regulatory Integrative Comp. Physiol 30):R1276-R1285, 1991; McKenna, etal., Pharm. Bioch. Behav., 40:151-156, 1991; and Takahashi, et al.,Brain Res., 359:194-207, 1985.

[0713] Preparation of the Compounds of the Invention

[0714] Preparation of the compounds of the present invention may becarried out via sequential or convergent synthetic routes. The skilledartisan will recognize that, in general, the three domains of a compoundof formula I are connected via amide bonds. The B and C domains areoptionally connected via a reduced or partially reduced amide bond(e.g., via reductive amination). The skilled artisan can, therefore,readily envision numerous routes and methods of connecting the threedomains via standard peptide coupling reaction conditions.

[0715] The phrase “standard peptide coupling reaction conditions” meanscoupling a carboxylic acid with an amine using an acid activating agentsuch as EDC, dicyclohexylcarbodiimide, andbenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphatein a inert solvent such as DCM in the presence of a catalyst such asHOBT. The uses of protective groups for amine and carboxylic acids tofacilitate the desired reaction and minimize undesired reactions arewell documented. Conditions required to remove protecting groups whichmay be present can be found in Greene, et al., Protective Groups inOrganic Synthesis, John Wiley & Sons, Inc., New York, N.Y. 1991.

[0716] CBZ, Boc and FMOC protecting groups are used extensively in thesynthesis, and their removal conditions are well known to those skilledin the art. For example, removal of CBZ groups can he achieved bycatalytic hydrogenation with hydrogen in the presence of a noble metalor its oxide such as palladium on activated carbon in a protic solventsuch as ethanol. In cases where catalytic hydrogenation iscontraindicated by the presence of other potentially reactivefunctionality, removal of CBZ can also be achieved by treatment with asolution of hydrogen bromide in acetic acid, or by treatment with amixture of TFA and dimethylsulfide. Removal of Boc protecting groups iscarried out in a solvent such as methylene chloride, methanol or ethylacetate with a strong acid, such as TFA or HCl or hydrogen chloride gas.

[0717] The compounds of formula I, when exist as a diastereomericmixture, may be separated into diastereomeric pairs of enantiomers byfractional crystallization from a suitable solvent such as methanol,ethyl acetate or a mixture thereof. The pair of enantiomers thusobtained may be separated into individual stereoisomers by conventionalmeans by using an optically active acid as a resolving agent.Alternatively, any enantiomer of a compound of the formula I may beobtained by stereospecific synthesis using optically pure startingmaterials or reagents of known configuration.

[0718] The compounds of the present invention can be prepared accordingto the procedure of the following schemes and examples, which mayfurther illustrate details for the preparation of the compounds of thepresent invention. The compounds illustrated in the examples are,however, not to be construed as forming the only genus that isconsidered as the present invention.

[0719] In the Schemes, Preparations and Examples below, various reagentsymbols and abbreviations have the following meanings: BINAP2,2’-Bis(diphenylphosphino)-1,1’-binaphthyl Boc t-butoxycarbonyl CBZbenzyloxycarbonyl DCM dichloromethane DEAD diethyl azodicarboxylate DIADdiisopropyl azodicarboxylate DIPEA diisopropylethylamine DMAP4-dimethylamino pyridine DMF N,N-dimethylformamide DMSOdimethylsulfoxide eq. equivalent(s) EDC1-(3-dimethylaminopropyl)-3-ethylcarbodiimide HCl ESI-MS electron sprayion-mass spectroscopy Et ethyl EtOAc ethyl acetate FMOC9-Flurorenylmethyl carbamate HATUO-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluroniumhexafluorophosphate HOAT 1-hydroxy-7-azabenzotriazole HOBT1-hydroxybenzotriazole hydrate HPLC high performance liquidchromatography HRMS high resolution mass spectroscopy h (hr) hour(s)LRMS low resolution mass spectroscopy Me methyl Ms methanesulfonyl NMM4-methyl morpholine Pd₂(dba)₃ tris(dibenzylideneacetone) dipalladium(0)Ph phenyl Phe phenylalanine Pr propyl r.t. room temperature TBAFtetrabutylammonium fluoride TBS tertbutyldimethylsilyl TFAtrifluoroacetic acid TEA triethylamine THF tetrahydrofuran Tic1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid TLC thin-layerchromatography

[0720] Reaction Scheme 1: Coupling Procedures

[0721] In coupling procedure 1, an appropriate A domain (e.g.,piperazine) is coupled to B domain (e.g., D-Boc-p-Cl-Phe-OH) in thepresence of EDC/HOBt followed by Boc deprotection. The coupled ABcompound is then coupled to an appropriate C domain followed bydeprotection of Boc group and salt formation. Alternatively, when Cdomain is not protected with Boc group, the final compound can beobtained without the deprotection step.

[0722] In coupling procedure 2, an appropriate A domain (e.g.,piperazine) is coupled to an appropriate BC domain in the presence ofHATU followed by deprotection of Boc group and salt formation.Alternatively, when BC domain is not protected with Boc group, the finalcompound can be obtained without the deprotection step.

[0723] In coupling procedure 3, an appropriate AB domain is coupled toan appropriate C domain in the presence of EDC/HOBt followed bydeprotection of Boc group and salt formation.

[0724] In coupling procedure 4, an appropriate BC domain is coupled toan appropriate A domain in the presence of EDC/HOBT followed bydeprotection of Boc group and salt formation. Alternatively, when Cdomain is not protected with Boc group, the final compound can beobtained without the deprotection step.

[0725] In coupling procedure 5, an appropriate AB domain is coupled toan appropriate C domain in the presence of HATU followed by deprotectionof Boc group salt formation.

[0726] For coupling of A with Boc-B, EDC/HOAT, EDC/HOBT or DCC/HOBT canbe used.

[0727] Generally, the starting material of Boc-protected piperazine (Adomain) can be deprotected in the presence of TFA/CH₂Cl₂, HCl/EtOAc,HCl/dioxane, or HCl in MeOH/Et₂O with or without a cation scavenger,such as dimethyl sulfide (DMS) before being subjected to the couplingprocedure. It can be freebased before being subjected to the couplingprocedure or in some cases used as the salt.

[0728] A suitable solvent such as CH₂Cl₂, DMF, THF or a mixture of theabove solvents can be used for the coupling procedure. Suitable baseincludes triethyl amine (TEA), diisopropyethyl amine (DIPEA),N-methymorpholine, collidine, or 2,6-lutidine. Base may not be neededwhen EDC/HOBt is used.

[0729] Generally after the reaction is completed, the reaction mixturecan be diluted with an appropriate organic solvent, such as EtOAc,CH₂Cl₂, or Et₂O, which is then washed with aqueous solutions, such aswater, HCl, NaHSO₄, bicarbonate, NaH₂PO₄, phosphate buffer (pH 7), brineor any combination thereof. The reaction mixture can be concentrated andthen be partitioned between an appropriate organic solvent and anaqueous solution. The reaction mixture can be concentrated and subjectedto chromatography without aqueous workup.

[0730] Protecting group such as Boc or CBZ, FMOC, CF₃CO and H₂/Pd—C canbe deprotected in the presence of TFA/CH₂Cl₂, HCl/EtOAc, HCl/dioxane,HCl in MeOH/Et₂O, NH3/MeOH, or TBAF with or without a cation scavenger,such as thioanisole, ethane thiol and dimethyl sulfide (DMS). Thedeprotected amines can be used as the resulting salt or are freebased bydissolving in CH₂Cl₂ and washing with aqueous bicarbonate or aqueousNaOH. The deprotected amines can also be freebased by ion exchangechromatography.

[0731] The compounds of the present invention can be prepared as salt,such as TFA, hydrochloride or succinate salts by using known standardmethods.

[0732] Reaction Scheme for Preparation of “A Domain”

[0733] The A domains of the present invention, in general, may beprepared from commercially available starting materials via knownchemical transformations. The preparation of A domain of the compound ofthe present invention is illustrated in the reaction scheme below.

[0734] Reaction Schemes of “A Domain”

[0735] Reaction Scheme 2: Buchwald

[0736] As shown in Reaction Scheme 2, the “A domain” of the compounds ofthe present invention can be prepared by coupling halo-substituted aryl1 (X-Q) with piperazines 2 in the presence of tris(dibenzylideneacetone)dipalladium (Pd₂(dba)₃), 1,1′-Bi[(2-diphenylphosphines) naphthalene](BINAP) and sodium t-butoxide (NaO^(t)Bu) or cesium carbonate (Cs₂CO₃)in an organic solvent such as toluene at a suitable temperature. Moredetailed examples of A Domain preparation are described below.

[0737] Reaction Scheme 3: SNAr

[0738] As shown in Reaction Scheme 3, the “A domain” of the compounds ofthe present invention can be prepared by heating appropriatelysubstituted fluoro-aryl compounds 4 and piperazines 2 neat or with anappropriate solvent and with or without an appropriate base.

[0739] Reaction Scheme 4: SNAr Followed by Buchwald

[0740] As shown in Reaction Scheme 4, the “A domain” of the compounds ofthe present invention can be prepared by heating1-bromo-2-fluoro-benzene 6 with various alcohols (R⁹-OH) in the presenceof NaH to give ortho-substituted bromobenzenes 7 which can then besubjected to Buchwald conditions as shown in Reaction Scheme 4 above.

[0741] Reaction Scheme 5:

[0742] Copper mediated O-arylation of 2-bromophenol with aryl boronicacids followed by Buchwald.

[0743] A is aryl or heteroaryl.

[0744] As shown in Reaction Scheme 5, the “A domain” of the compounds ofthe present invention can be prepared by heating 2-bromophenol 9 withvarious aryl and heteroaryl boronates (X-OH) in the presence of Cu(OAc)₂and pyridine to give ortho-substituted bromobenzenes 10 which can thenbe subjected to Buchwald conditions.

[0745] Reaction Scheme 6: Benzylamines

[0746] 6A. Nitrile Reduction

[0747] A=SO₂R⁹, SO₂N(R⁹)₂, C(O)R⁹, C(O)OR⁹, C(O)SR⁹, C(O)N(R⁹)₂ and etc.

[0748] As shown in Reaction Scheme 6A, the “A domain” of the compoundsof present invention can be prepared by reducing the nitrile of(2-cyano-phenyl)-piperazine 12 to the corresponding benzyl amine 13 witheither NaBH₄ and TFA or H₂ and Raney nickel. Benzyl amine 13 can betransformed to other benzyl amine derivatives 14 using various methodsknown to the skilled artisan.

[0749] 6B. From benzylic alcohol via Mitsunobu or via mesylate

[0750] A=acidic heteroaryl, azide, imide and etc.

[0751] B=basic heteroaryl, heterocyclyl and etc.

[0752] As shown in Reaction Scheme 6B, the “A domain” of the compoundsof present invention can be prepared by hydrolyzing the nitrile of(2-cyano-phenyl)-piperazine 12 to the corresponding carboxylic acid 15with KOH followed by reduction to benzyl alcohol 16 with BH₃-ThF. Benzylalcohol 16 can be transformed to benzyl amines 17 either using Mitsunobuconditions or by activating the alcohol as the mesylate followed bynucleophilic displacement.

[0753] Reaction Scheme 7: Derivatives of1-Boc-4-(2-amino-phenyl)-piperazine

[0754] A=SO₂R⁹, SO₂N(R⁹)₂, C(O)R⁹, C(O)OR⁹, C(O)SR⁹, C(O)N(R⁹)₂ and etc.

[0755] As shown in Reaction Scheme 7, the “A domain” of the compounds ofpresent invention can be prepared from1-Boc-4-(2-amino-phenyl)-piperazine 19 which is prepared from4-(2-nitro-phenyl)-piperazine 18 by Boc protection followed by nitroreduction. 1-Boc-4-(2-amino-phenyl)-piperazine 19 can be transformed toother aniline derivatives 20 using various methods known to the skilledartisan. Sulfonamides 21 could be prepared from1-Boc-4-(2-amino-phenyl)-piperazine 19 by reaction with various sulfonylchlorides. The resulting sulfonamides 21 could then be deprotonated withNaH or K₂CO₃ in DMF followed by alkylation with various alkyl halides(R⁹X) to afford alkylated sulfonamides 22.1-Boc-4-(2-amino-phenyl)-piperazine could also be acylated with variousacid chlorides to give acetamides 23. The acetamides 23 could be reducedwith BH₃-THF to give alkyl amines 24 which can be transformed to otheramine derivatives 25 using various methods known to the skilled artisan.

[0756] Reaction Scheme 8: Derivatives of2-(N-Boc-piperazin-1-yl)-benzaldehyde

[0757] As shown in Reaction Scheme 8, the “A domain” of the compounds ofpresent invention can be prepared by reducing the nitrile of(2-cyano-phenyl)-piperazine 12 to the corresponding aldehyde 26 withDIBAL. Aldehyde 26 can be transformed to benzyl amines 27 by reductiveamination with various amines including nitrogen containingheterocycles. These benzyl amines 27 can be transformed to other aminederivatives using various methods known to the skilled artisan.

[0758] Aldehyde 26 can also be reacted with various organolithiumreagents (including lithiated aryl and heteroaryl groups) to givealcohols 28. The alcohol can be oxidized to give ketones 29 or removedby Barton deoxygenation to give 30.

[0759] Reaction Scheme 9: Derivatives of1-Boc-4-(2-hydroxy-phenyl)-piperazine

[0760] As shown in Reaction Scheme 9, “A domain” of the compounds ofpresent invention can be prepared by treating1-Boc-(2-hydroxy-phenyl)-piperazine 31 with a base and an alkyl halide(RX) or subjected to Mitsunobu conditions with R⁹OH to giveortho-substituted aryl piperazines 32.

[0761] Reaction Scheme 10: Derivatives of1-Boc-4-(2-carboxy-phenyl)-piperazine

[0762] A=heterocyclyl, N(R⁹)₂, OR⁹ or SR⁹ and etc.

[0763] As shown in Reaction Scheme 10, “A domain” of the compounds ofpresent invention can be prepared by hydrolyzing the nitrite of(2-cyano-phenyl)-piperazine 12 to the corresponding carboxylic acid 15with KOH followed by transformation to other carboxylic acid derivatives33 using various methods known to the skilled artisan.

[0764] Reaction Scheme 11: Tetrazoles

[0765] As shown in Reaction Scheme 11, “A domain” of the compounds ofpresent invention can be prepared by reacting the nitrile of(2-cyano-phenyl)-piperazine 12 with tributyltin azide to give tetrazoles34. The tetrazoles can be further transformed to 35 using variousmethods known to the skilled artisan.

[0766] The present invention also provides a novel process for preparingcertain intermediates and/or compounds of the invention as shown inReaction Schemes 12-14.

[0767] Reaction Scheme 12:

[0768] AS shown in Reaction Scheme 12, a convergent synthesis of a keyintermediate isoindoline (5) via a Heck coupling, followed by areductive amination, a ring cyclization and a resolution has beendeveloped. Also, alternate asymmetric approaches including asymmetricMichael addition and asymmetric hydrogenation have also been developedto prepare compounds of the invention and/or intermediates thereof.

[0769] As shown in Reaction Scheme 12, the isoindoline compounds of thepresent invention may be prepared from 2-halobenzaldehyde 1 orsubstituted analog thereof. Preferred starting material is2-bromobenzaldehyde or substituted analog thereof. Pd-mediated Heckcoupling of 2-bromobenzaldehydes 1 with for example, methyl acrylate,provided alpha, beta-unsaturated methyl esters 2, which undergoesreductive amination to give amines, 3 (or carbamates where R₁ is forexample, Boc). Various Heck coupling reagents and conditions were foundsuitable to effect the coupling reaction. Suitable catalysts and ligandsinclude Pd(OAc)₂/PPh₃, Pd(OAc)PPh₃/BU₄NBr, Pd(PPH₃)₂Cl₂/CUI,Pd(OAC)₂/P(O-Tol)₃. Suitable solvent or solvent systems for the Heckcoupling reaction include DMF, toluene and ethyl acetate. More preferredbase is triethylamine.

[0770] Reductive amination of the aldehyde functionality of 2 to aminesis accomplished in good yields by reaction with benzylamine oralpha-methylbenzylamine in acidic conditions, followed by in situreduction of the incipient imines with NaCNBH₃ at about pH 5. Otherreducing agents including Na(OAc)₃BH and NaBH₄/H may also be used toeffect reduction of the incipient imines. Interestingly, the resultingamines immediately cyclized to the isoindoline compounds under the sameacidic conditions for the reduction. Direct preparation of compound 4may also be effected by use of BocNH₂ instead of benzylamine in thereductive amination step. Screening of various reducing agentsdemonstrated that the combination of Et₃SiH and TFA in CH₃CN representsthe preferred method for effecting reductive amination using BocNH₂.

[0771] The N-Boc isoindolinecarboxylic acid 5 may also be prepared from3 as the carbamate, by an intra-molecular Michael addition and esterhydrolysis. The resolution of the isoindolinecarboxylic acids 4 bycrystallization afforded enantio-pure compounds 5.

[0772] Two alternate asymmetric approaches have also been developed forthe synthesis of isoindolinecarboxylic acid 5 i.e., asymmetric Michaeladditions and asymmetric hydrogenation. In the asymmetric Michaeladdition approach, alpha-methylbenzyl amine is used as a chiralauxiliary to induce the enantio-selectivity. In the asymmetrichydrogenation approach, compound 4′ could be converted to 5stereoselectively in the presence of chiral ligands.

[0773] Finally the coupling of the isoindolines 5 with the “B” domainpiece, i.e., D-Cl-Phe to afford compound 6 (“BC” piece) is accomplishedby standard amino acid coupling reactions such as, for example, by theuse of EDC or EDCI or other activating agents in the presence ofsuitable is dimethylaminopyridine (DMAP). The product (6) is thencoupled with an “A” domain piece to afford the target MC4R agonistcompound of formula I by coupling reactions known to one of skill in theart.

[0774] Preferably, the isoindole or other “C” domain piece is coupled toan “AB” coupled domain piece to form the compound of formula I.

[0775] Reaction Scheme 13:

[0776] As shown in Reaction Scheme 13, m-tyrosine ester or analogs,including substituted analogs thereof, may be esterified by forming theacid halide followed by nucleophilic displacement of halide by thealkoxy group from an alcohol, i.e., methanol or ethanol. Where thionylchloride or other halide source is used the product may be isolated asthe acid addition salt (2). The resulting ester (2) is subjected to aPictet-Spengler reaction by heating with a suitable ketone or aldehydein refluxing conditions. For example, an unsubstituted isoquinolinebackbone (3) may be formed by employing formaldehyde in thepictet-Spengler reaction. On the other hand, a gem-dimethyl substitutedisoquinoline wherein R¹¹ is methyl, may be formed by using acetone asthe ketone source and solvent. Other less reactive substituents may besubstituted as the R¹¹ group for the practice of the present invention.

[0777] The product isoquinoline (3) may be isolated preferably as theacid addition salt. Where m-tyrosine is used as the starting material,the free hydroxyl group is removed first by protection/activation with agood leaving group such as, for example, reaction with triflic anhydride(trifluoromethane sulfonic anhydride) or methanesulfonic acid to formthe triflate or mesylate in the presence of a base. The triflate is apreferred group used to set up the compound (3) for deoxygenationbecause of the extra electron withdrawing effect of the trifluoromethanesubstituent. The deoxygenation reaction is effected by hydrogenation atpressures of about 50 psi. The product (4) may be isolated as the acidaddition salt. The product (4) is hydrolyzed under basic conditions toafford the acid salt. Suitable bases for the above hydrolysis includeaqueous sodium hydroxide, potassium hydroxide and sodium lithiumhydroxide. The reaction is preferably performed in a mixture of aqueousand organic solvents. An exotherm during addition of base may beregulated (i.e., less than about 35° C.) to avoid overheating or“runaway reactions.” The reaction product may be isolated by aqueouswork up. Alternatively, the entire mixture may be concentrated andwashed with organic solvents to afford the desired product (6) aftercrystallization.

[0778] The product (6) is then reacted with a “B” domain substrate suchas, for example, 4-chloro-D-phenylalanine as described previously and inthe experimental section. The resulting “BC” combination product is thenreacted with an “A” domain piece to form the respective compound offormula I. Alternatively, the product (6) may be reacted with an “AB”domain combination product to afford a compound of formula I.

[0779] One of skill is aware that certain protections and deprotectionsof intermediates in Reaction Scheme 13, to form the carbamate,substituted amine or free amine at the isoquinolinyl nitrogen arepossible and contemplated as within the scope of this invention. Unlessotherwise specified, reagents and procedures for effecting the reactionsdescribed herein are known to one of skill in the art and may be foundin general reference texts such as Advanced Organic Chemistry by J.March, 5^(th) edition, Wiley Interscience Publishers, New York, N.Y.,and references therein.

[0780] In an alternate procedure, the isoquinoline product i.e.,compound (3) or (5) including their N-protected analogs may be resolvedby reaction with a resolving agent such as for example, L-tartaric acid,dehydroabietylamine or other resolving agents known to one of skill inthe art.

[0781] Alternatively, asymmetric analogs of product (6) may be preparedby using asymmetric starting materials. For example, L-DOPA may be usedin place of m-tyrosine ester in reactions essentially similar to thosedescribed and illustrated in Reaction Scheme 13, and in the examples, toafford the asymmetric analog of compound (6).

[0782] Tetrahydroisoquinoline acetic acid derivatives may be preparedand utilized as shown in Reaction Scheme 14 below:

[0783] Reaction Scheme 14:

[0784] As shown in Reaction Scheme 14, a compound of formula 10a whereinX is halogen, preferably bromo or chloro, and R and R¹¹ are as definedpreviously, and which is obtained commercially or prepared fromcommercial starting materials is reacted with cyanomethylethylacetate toafford a compound of formula 10b. The compound of formula 10b may beprotected as the compound 10c with a suitable protecting group (Pg) andthen subjected to hydrogenation conditions including for exampleasymmetric hydrogenation to form a compound of formula 10d, which may bechiral (depending on hydrogenation conditions, i.e., asymmetric versusnon-asymmetric hydrogenation). The compound of formula 10d orstereoisomer thereof, is reacted with a B-domain piece such as, forexample, 4-chloro-D-phe to afford a BC piece (10e). The compound offormula 10e is then reacted with an A-domain piece to afford a compoundof formula I. The details of the specific reaction steps are similar toor analogous to reactions taught herein, and in the experimentalsection. Furthermore, one of skill in the art is aware of that suchintermediate reactions as hydrolysis and deprotection may be necessaryto achieve optimum yields in certain steps of the scheme as shown. Oneof skill in the art is also aware of further common manipulations suchas N-alkylation, or N-acylation, and alkylations on the benzene ring toafford other compounds of formula I.

[0785] The following describes the detailed examples of A Domainpreparation.

Preparation 1A (Buchwald Using NaOtBu)(3R)-3-methyl-(2-methylthiophenyl)piperazine

[0786]

[0787] 2-Bromothioanisole (300 mg, 1.48 mmol), (R)-2-methylpiperazine(185 mg, 1.85 mmol), Pd₂(dba)₃ (32 mg, 0.35 mmol), BINAP (41 mg, 0.66mmol), sodium t-butoxide (200 mg, 2.08 mmol) and anhydrous toluene (3ml) were combined in a 15 ml round-bottomed flask. The atmosphere in theflask was evacuated and flushed with nitrogen (3×). The mixture waslowered into an oil bath heated to 100° C. After heating for about 1.2hours, the mixture was cooled, diluted with ethyl acetate (100 ml),filtered through Celite, and concentrated to a crude oil (285 mg). Theoil was loaded onto an cation exchange column, and the column wasflushed with methanol (100 ml), and then with 2 M ammonia/methanol (100ml). The basic methanol solution was concentrated to an oil (250 mg).The oil was further purified by flash chromatography using 19:1dichloromethane: 0.5 M ammonia/methanol as eluent to yield the finalproduct (160 mg, 58%) as an oil. LRMS (ESI+): 223.0 (M+1)

Preparation 2A (Buchwald Using CsCO₃)4-(2-diethylcarbamoyl-phenyl)-piperazine

[0788]

[0789] HOBT (2.72 g, 10.08 mmol), DIPEA (3.52 mL, 20.16 mmol), 2-bromobenzoic acid (4.08 g, 10.08 mmol), and diethyl amine (2.08 mL, 10.08mmol) were dissolved in DCM (100 mL) and stirred at r.t. for about 30minutes. EDCI (3.86 g, 10.08 mmol) was added, and the mixture wasstirred at r.t. for about 16 hours. The reaction was concentrated to anoil, and the oil was purified via column chromatography to give2-bromo-N,N-diethyl-benzamide (3.35 g, 68%) as a yellow oil.

[0790] Piperazine (489 mg, 4.8 mmol), 2-bromo-N,N-diethyl-benzamide (1g, 3.95 mmol), Pd₂(dba)₃ (235 mg, 0.2 mmol), BINAP (442 mg, 0.6 mmol),and cesium carbonate (3 g, 5.53 mmol) were mixed together in toluene (20mL). The mixture was degassed and heated to 100° C. for about 72 hours.The mixture was diluted with ether (100 mL) and filtered over celite.The filtrate was concentrated and then subjected to chromatography onsilica gel to give the title compound (480 mg, 47%) as a brown oil. LRMS(ESI+): 262.2 (M+1)

Preparation 3A 1-Boc-4-(2-piperazin-1-yl-benzoyl)-piperazine

[0791]

[0792] Boc protected piperazine (849 mg, 4.56 mmol) was dissolved in DCM(20 mL) and triethyl amine (2.54 mL, 18.2 mmol) was added. To thestirred solution, ortho-bromo benzoyl chloride (2 g, 9.11 mmol) wasadded via syringe under nitrogen. The system was stirred for about 12hours at r.t. The reaction was washed with water, dried, filtered, andconcentrated. The residue was subjected to chromatography on silica gelto give 1-Boc-4-(2-bromo-)-piperazine (1.48 g, 8.85 mmol) as a whitefoam. 1-Boc-4-(2-bromo-benzoyl)-piperazine was coupled to piperazine ina manner similar to Preparation 1A. LRMS (ESI+): 375.2 (M+1)

Preparation 4A 1-(2-Methoxy-5-nitro-phenyl)-piperazine

[0793]

[0794] 1-(2-Methoxy-5-nitro-phenyl]piperazine was prepared in a mannersimilar to Preparation 1A except that piperazine was coupled to2-bromo-1-methoxy-4-nitro-benzene. LRMS (ESI+): 238.4 (M+1)

Preparation 5A 1-(2-Methyl-6-nitro-phenyl)-piperazine

[0795]

[0796] 1-(2-Methyl-6-nitrophenyl]piperazine was prepared in a mannersimilar to Preparation 1A except that piperazine was coupled to2-bromo-1-methyl-3-nitro-benzene. LRMS (ESI+): 222.4 (M+1)

Preparation 6A 1-(2-isopropoxy-phenyl)-piperazine

[0797]

[0798] The title compound was prepared in a manner similar toPreparation 1A except that piperazine was coupled to1-bromo-2-isopropoxy-benzene. LRMS (ESI+): 221.4 (M+1)

Preparation 7A 1-(2-isopropyl-phenyl)piperazine

[0799]

[0800] The title compound was prepared in a manner similar toPreparation 1A except that piperazine was coupled to1-bromo-2-isopropyl-benzene. LRMS (ESI+): 205.4 (M+1)

Preparation 8A 1-(2-isopropyl-5-methyl-phenyl)piperazine

[0801]

[0802] The title compound was prepared in a manner similar toPreparation 1A except that piperazine was coupled to1-bromo-5-methyl-2-isopropyl-benzene.

[0803]¹H NMR (CDCl₃) δ7.05-7.00 (m, 1H), 6.85-6.75 (m, 2H), 3.95 (s,1H), 3.10-3.00 (m, 4H), 2.95-2.90 (m, 4H) 2.30 (s, 3H,), 1.25-1.20 (m,6H).

Preparation 9A 1-(2-cyclohexyl-phenyl]piperazine

[0804]

[0805] The title compound was prepared in a manner similar toPreparation 1A except that piperazine was coupled to1-bromo-2-cyclohexyl-benzene. LRMS (ESI+): 245.1 (M+1)

Preparation 10A 1-[2-(1,1-Difluoro-ethyl)-phenyl]-piperazine

[0806]

[0807] A solution of diethylaminosulfur trifluoride (560 mg, 3.47 mmol,3 eq) and 2-bromoacetophenone (230 mg, 1.16 mmol, 1.0 eq) was heated to40° C. for about 72 hours. The solution was diluted with CH₂Cl₂ andwashed with saturated sodium bicarbonate, water, brine, dried overNa₂SO₄, filtered, and concentrated. Purification by flash chromatography(35 g SiO₂, linear gradient 0-10% ethyl acetate/Hexanes, 30 mL/minute,over 30 minutes) afforded about 125 mg (0.57 mmol, 49%) of2-(1,1-difluoroethyl)-1-bromobenzene. GC/MS (EI): 220 (M+H).2-(1,1-Difluoroethyl)-1-bromobenzene was coupled to piperazine in amanner similar to Preparation 1A. LRMS (ESI+): 227.2 (M+1)

Preparation 11A(S)-1-{2-[1-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-phenyl}-piperazine

[0808]

[0809] To a 25 mL flask containing (S)-(−)-2-bromo-alpha-methylbenzylalcohol (200 mg, 1.0 mmol), tert-butyldimethylsilyl chloride (165 mg,1.1 mmol), and imidazole (203 mg, 3.0 mmol) flushed with nitrogen wasadded 5 mL of dimethylformamide. After stirring overnight, the mixturewas quenched with saturated sodium bicarbonate, diluted with ethylacetate, washed with NaH₂PO₄, saturated aqueous sodium bicarbonate,water, brine, dried (Na₂SO₄), filtered, and concentrated. Purificationby flash chromatography (10 g SiO₂, linear gradient 0-10% ethylacetate/Hexanes, 30 mL/minute, over 30 minutes) gave about 260 mg (0.82mmol, 82%) of (5)-[1-(2-Bromo-phenyl)-ethoxy]-tert-butyl-dimethyl-silaneas a colorless oil. GC/MS (EI): 315 (M). The(5)-[1-(2-Bromo-phenyl)-ethoxy]-tert-butyl-dimethyl-silane was coupledto piperazine in a manner similar to Preparation 1A. LRMS (ESI+): 321.5(M+1)

Preparation 12A(R)-1-{2-[1-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-phenyl}-piperazine

[0810]

[0811] The title compound was prepared in a manner similar toPreparation 11A except that (R)-(−)-2-bromo-alpha-methylbenzyl alcoholwas used. LRMS (ESI+): 321.3 (M+1)

Preparation 13A (2R)-3-Ethyl-1-(2-methylthiophenyl)piperazine

[0812]

[0813] The title compound was prepared in a manner similar toPreparation 1A. LRMS (ESI+): 237.1 (M+1)

Preparation 14A (3S)-3-Methyl-1-(2-methylthiophenyl)piperazine

[0814]

[0815] The title compound was prepared in a manner similar toPreparation 1A.

Preparation 15A 1-(2-Ethylphenyl)piperazine

[0816]

[0817] The title compound was prepared in a manner similar toPreparation 1A. LRMS (ESI+): 191.2 (M+1)

Preparation 16A (2R)-2-methyl-1-(2-methylthophenyl)piperazine

[0818]

[0819] (2R)-4-Benzyl-2-methyl-1-(2-methylthiophenyl) piperazine wasprepared in 26% yield from ortho-bromothioanisole and(R)-3-methyl-1-benzylpiperazine in a manner similar to Preparation 1A.LRMS (ESI+): 223.2 (M+1)

[0820] (2R)-4-Benzyl-2-methyl-1-(2-thiomethylphenyl) piperazine (24 mg,0.077 mmol) was dissolved in 1,2-dichloroethane (4 ml) and cooled in anice bath. To the chilled solution was added 1-chloroethyl chloroformate(38 microliters, 50 mg, 0.35 mmol) in one portion. The solution wascovered with a nitrogen atmosphere and then heated to 50° C. Afterstirring at 50° C. for about 1.25 hours, the solution was concentratedunder reduced pressure and then dissolved in methanol (6 ml). Themethanolic solution was covered with a nitrogen atmosphere and allowedto stir overnight at r.t. The solution was concentrated to give about 21mg of crude oil. Flash chromatography (10% 0.5 M NH₃/methanol in DCM aseluent) yielded the final compound (14 mg, 82%). LRMS (ESI+): 223.2(M+1)

Preparation 17A (2S)-2-Methyl-1-(2-methylthiophenyl)piperazine

[0821]

[0822] The title compound was prepared in a manner similar toPreparation 16A. LRMS (ESI+): 223.2 (M+1)

Preparation 18A 1-[2-(2-Methyl-propane-1-sulfonyl)-phenyl]-piperazine

[0823]

[0824] To a solution of 2-bromobenzene thiol (10.0 g, 52.8 mmol, 1.0eq.) in DMF (250 mL) was added K₂CO₃ (17.5 g, 126.7 mmol, 2.4 eq.) andisobutyl iodide (7.3 mL, 63.36 mmol, 1.2 eq). The reaction was warmed toabout 40° C. and stirred overnight. The mixture was diluted with EtOAc(300 mL) and washed with water (100 mL) and brine (100 mL). The organiclayer was extracted with EtOAc(2×). The combined organic extracts weredried (Na₂SO₄), filtered and concentrated to afford1-bromo-2-isobutylsulfanyl-benzene (12.94 g, 52.8 mmol, 100%), which isthen taken to the next step without any further purification. GCMS (EI):244.0

[0825] To a solution of 1-bromo-2-isobutylsulfanyl-benzene (8.0 g, 32.6mmol, 1.0 eq) in DCM (100 mL) at 0° C. was added CaCO₃ (13.05 g, 130.4mmol, 4.0 eq.) and MCPBA (28.1 g, 81.5 mmol, 2.5 eq.). The mixture wasstirred for about 30 minutes and filtered through a pad of celite. Thesolution was washed with sodium bisulfite (2×) and 5N NaOH (2×). Theorganic layer was dried (Na₂SO₄), filtered and concentrated.Purification by flash chromatography (250 g SiO₂, linear gradient, 40mL/min, 10%-40% EtOAC/hexane for about 33 minutes) afforded1-bromo-2-(2-methyl-propane-1-sulfonyl)-benzene (7.4 g, 26.6 mmol, 82%).GCMS (EI): 276.0. 1-bromo-2-(2-methyl-propane-1-sulfonyl)-benzene wascoupled to piperazine in a manner similar to Preparation 1A. LRMS(ESI+): 283.06 (M+1)

Preparation 19A (SNAr) 1-(2-aminosulfonyl-phenyl)piperazine

[0826]

[0827] To a 50 mL flask containing 2-flourobenzene sulfonamide (200 mg,1.14 mmol, 1 eq.) and piperazine (245 mg, 2.84 mmol, 2.5 eq) was added20 mL of dioxane. The solution was heated to 100° C. for about 4 hours.More piperazine (200 mg, 2.32 mmol, 2 eq.) was added and the solutionwas heated to 100° C. for another 72 hours. The solution wasconcentrated to an oil and dissolved in 30 mL of 0.1 M pH 7.0 phosphatebuffer. The aqueous solution was extracted with CH₂Cl₂ (3×30 mL). Thecombined organic extracts were dried over Na₂SO₄ and concentrated toyield about 275 mg (1.14 mmol, 100%) of the title compound. LRMS (ESI+):242.1 (M+H).

Preparation 20A 1-Boc-4-(3-Chloro-2-cyano-phenyl)-piperazine

[0828]

[0829] To a solution of N-Boc-piperazine (2.02 g, 11.0 mmol) in DMSO (20mL) was added 2-fluoro-6-chlorobenzonitrile (1.55 g, 10 mmol) andpotassium carbonate (1.52 g, 11 mmol). The mixture was stirred at 80° C.for about 48 hours. The mixture was cooled to r.t. and diluted withdiethyl ether (200 mL). The solution was washed with iN HCl (2×20 mL),H₂O (3×20 mL) and brine (20 mL) and then dried over sodium sulfate andconcentrated to a yellow oil. Purification by flash chromatography (4:1hexanes/ethyl acetate) gave the title compound (2.5 g, 86%) as acolorless oil.

[0830]¹H NMR (CDCl₃) δ7.40-7.50 (m, 1H,), 7.10-7.20 (m, 1H), 6.80-6.90(m, 1H), 3.70 (s, 4H), 3.20 (s, 4H), 1.48 (s, 9H). TLC (SiO₂): 0.48 (4:1hexanes/ethyl acetate)

Preparation 21A1-Boc-4-(3-Chloro-2-dimethylaminomethyl-phenyl)-piperazine

[0831]

[0832] Sodium borohydride (1.2 g, 31.4 mmol) was dissolved in THF (20mL) and TFA (2.42 mL, 31.4 mmol) in THF (20 mL) was added dropwise at 0°C., and the reaction was stirred for about 30 minutes.1-Boc-4-(3-Chloro-2-cyano-phenyl)-piperazine (2.0 g, 6.3 mmol) wasdissolved in ThF (20 mL) and added dropwise to the solution at 0° C.,and the reaction was stirred for about 24 hours. The reaction wascarefully quenched with H₂O and ethyl acetate (200 mL) was added. Themixture was washed with H₂O (3×25 mL), brine (25 mL) and dried overMgSO₄. The solvents were removed in vacuo and the crude reaction mixturewas dissolved into acetonitrile (7 mL). Formalin (1.6 mL, 59.2 mmol) wasadded, followed by sodium cyanoborohydride (0.26 g, 7.4 mmol) at 0° C.The reaction was warned to r.t. and stirred for about one hour. Thereaction was quenched with H₂O and ethyl acetate (100 mL) was added. Thesolution was washed with saturated NaHCO₃ (2×10 mL) and dried overMgSO₄. Purification by silica gel chromatography (1:1 hexanes/ethylacetate) gave the title compound as a yellow oil (180 mg, 13%).

[0833]¹H NMR (CDCl₃) δ7.10-7.15 (m, 2H,), 6.92-6.98 (m, 1H), 3.68 (s,2H), 3.50-3.60 (m, 4H), 2.90-2.97 (m, 4H), 2.25 (s, 6H), 1.48 (s, 9H).TLC (SiO₂): 0.28 (1:1 hexanes/ethyl acetate)

Preparation 22A 1-Boc-4-(2-cyano-phenyl)-[1,4]diazepane

[0834]

[0835] To a solution of 1-Boc-homopiperazine(2.18 g, 11.0 mmol) in DMSO(20 mL) was added 2-fluorobenzonitrile (1.21 g, 1.08 mL, 10 mmol) andpotassium carbonate (1.52 g, 11 mmol). The mixture was stirred at 80° C.for about 48 hours. The mixture was cooled to r.t. and diluted withdiethyl ether (200 mL). The solution was washed with 1N HCl (2×20 mL),H₂O (3×20 mL) and brine (20 mL), dried over sodium sulfate andconcentrated to a yellow oil. Purification by flash chromatography (3:1hexanes/ethyl acetate) gave the title compound (1.1 g, 36%) as acolorless oil.

[0836]¹H NMR (CDCl₃) δ7.49 (dd, J=6.7, 1.7 Hz, 1H), 7.38 (td, J=7.3, 1.7Hz, 1H), 6.92 (d, J=8.5 Hz, 1H), 6.83 (t, J=7.4 Hz, 1H), 3.63-3.66 (m,2H), 3.46-3.57 (m, 6H), 2.01-2.10 (m, 2H), 1.40-1.45 (m, 9H). TLC(SiO₂): R_(f)=0.38 (3:1 hexanes/ethyl acetate)

Preparation 23A 1-Boc-4-(2-dimethylaminomethyl-phenyl)-[1,4]diazepane

[0837]

[0838] A solution of 1-Boc-4-(2-Cyano-phenyl)-[1,4]diazepane (600 mg,2.0 mmol) and Raney nickel (50% dispersion in H₂O, 1 mL) in methanol (50mL) was stirred under hydrogen (1 atm) for about 16 hours. Formalin (2mL) was added and the solution was stirred a further 24 hours. Themixture was filtered through celite. The filter cake was rinsed withmethanol (100 mL) and the filtrate was concentrated to a clear oil.Purification by flash chromatography (1% methanol/ethyl acetate) gavethe title compound (285 mg, 55%) as a colorless oil.

[0839]¹H NMR (CDCl₃) δ7.39 (d, J=7.4 Hz, 1H), 7.19 (t, J=7.4 Hz, 1H),7.03-7.10 (m, 2H), 3.53-64 (m, 4H), 3.52 (s, 2H), 3.04-3.08 (m, 4H),2.25 (s, 6H), 1.88-1.94 (m, 2H), 1.49 (s, 9H). TLC (SiO₂): R_(f)=0.40(ethyl acetate)

Preparation 24A (SNAr then Buchwald)1-(2-cyclohexyloxy-phenyl)-piperazine

[0840]

[0841] NaH (8.4 g, 210 mmol, 60% in mineral oil) was slurred in DMF (40mL) and heated to about 65° C. To the slurry was added cyclohexanol (7g, 69.9 mmol) dissolved in DMF (50 mL). The mixture was stirred at 65°C. for about 1 hour. Ortho-fluoro bromobenzene (9.2 mL, 83.9 mmol) wasadded dropwise in DMF (10 mL) and the mixture was stirred at 65° C. forabout 16 hours, and quenched with water and diluted with DCM. Themixture was concentrated to an oily solid and extracted between waterand 1/1 EtOAc/hex. The organic layer was dried, filtered andconcentrated. Chromatography on silica gel (EtOAc/hexanes) gave1-bromo-2-cyclohydroxy-benzene (6.13 g, 34%) as a yellow oil.1-Bromo-2-cyclohexyloxy-benzene was coupled to piperazine using theBuchwald chemistry described in preparation 1A. LRMS (ESI+): 261.1 (M+1)

Preparation 25A 1-(2-cycloheptyloxy-phenyl)-piperazine

[0842]

[0843] The title compound was prepared in a manner similar topreparation 24A except that cycloheptanol was used. LRMS (ESI+): 275.2(M+1)

Preparation 26A 1-[2-(3,3-dimethyl-cyclohexyloxy-phenyl)]-piperazine

[0844]

[0845] The title compound was prepared in a manner similar topreparation 24A except that 3,3-Dimethyl-cyclohexanol was used. LRMS(ESI+): 289.2 (M+1)

Preparation 27A 1-(2-cyclopentyloxy-phenyl)-piperazine

[0846]

[0847] The title compound was prepared in a manner similar topreparation 24A except that cyclopentanol was used. LRMS (ESI+): 247.1(M+1)

Preparation 28A 1-[2-(tetrahydro-thiopyran-3-yloxy)-phenyl]-piperazine

[0848]

[0849] The title compound was prepared in a manner similar topreparation 24A except that tetrahydro-thiopyran-3-ol was used. LRMS(ESI+): 279.2 (M+1)

Preparation 29A 1-[2-(tetrahydro-pyran-3-yloxy)-phenyl]-piperazine

[0850]

[0851] 3-hydroxy-tetrahydro pyrane was prepared according to Brown,Herbert C.; Prasad, J. V. N. Vara; Zee, Sheng-Hsu; J.Org.Chem. 50 (10),1985, 1582-1589. The compound was reacted with ortho-fluoro bromobenzenefollowed by Buchwald coupling in a manner similar to preparation 24A toafford the title compound. LRMS (ESI+): 263.1 (M+1)

Preparation 30A1-[2-(1,1-Dioxo-hexahydro-1λ⁶-thiopyran-4-yloxy)-phenyl]piperazine

[0852]

[0853] Ortho-fluoro bromobenzene was reacted withtetrahydro-thiopyran-3-ol to give4-(2-Bromo-phenoxy)-tetrahydro-thiopyran in a manner similar topreparation 24A. 4-(2-Bromo-phenoxy)-tetrahydro-thiopyran (1.94 g, 7.10mmol) is placed in DCM (70 mL) and calcium carbonate (2.84 g, 28.41mmol) was added. To this mixture cooled to 0° C. in an ice bath wasadded meta-chloro peroxy-benzoic acid (6.13 g, 17.75 mmol 50%) inportions while monitoring the temperature. The mixture was allowed towarm to r.t and stirred for about 15 minutes. The mixture was filteredover celite, and washed with sodium bisulfite solution (2×250 mL) andsodium bicarbonate (2×250 mL). The mixture was then concentrated to anoil. Chromatography (EtOAc/hexanes) provided4-(2-Bromo-phenoxy)-tetrahydro-thiopyran 1,1-dioxide (2.2 g, quant.) asa yellow solid. 4-(2-Bromo-phenoxy)-tetrahydro-thiopyran 1,1-dioxide wascoupled to piperazine using the Buchwald chemistry described inpreparation 1A to afford the title compound. LRMS (ESI+): 311.1 (M+1)

Preparation 31A (o-arylation of 2-bromophenol Followed by Buchwald)1-[2-(Pyridin-3-yloxy)-phenyl]-piperazine

[0854]

[0855] 2-Bromophenol (355 mg, 2.05 mmol), 3-pyridyl-boronic acid (500mg, 4.1 mmol), copper acetate (745 mg, 4.1 mmol) and pyridine (3.3 mL,41 mmol) were added to dichloromethane (41 mL) and stirred for about 48hours under air. The reaction was diluted with water (50 mL) and thelayers separated. The organic layer was washed with 5N NaOH. The organiclayer was concentrated, and chromatographed on silica gel(MeOH/dichloromethane) to yield 3-(2-bromo-phenoxy)-pyridine (30 mg, 6%)as a yellow oil. MS found 249.1 M+1. 3-(2-Bromo-phenoxy)-pyridine wascoupled to piperazine using the Buchwald chemistry described inpreparation 1A to afford the title compound. LRMS (ESI+): 256.1 (M+1)

Preparation 32A 1-(2-Phenoxy-phenyl)-piperazine

[0856]

[0857] A mixture of phenylboronic acid (5.12 g, 42 mmol), 2-bromophenol(3.55 g, 21 mmol), Cu(OAc)₂ (7.63 g, 42 mmol), pyridine (8 ml, 103 mmol)and 4 Å molecular sieves (2.1 g) in CH₂Cl₂ was stirred at r.t.overnight. The mixture was diluted with CH₂Cl₂, filtered through celite,washed with 1M NaOH, brine and dried. Removal of solvent gave1-bromo-2-phenoxybenzene, crystals (1.40 g, 27%). LRMS (ESI⁺): 248(M+1). 1-Bromo-2-phenoxybenzene was coupled to piperazine using theBuchwald chemistry described in preparation 1A to afford the titlecompound. LRMS (ESI+) 255 (M+1)

Preparation 33A 1-(2-m-tolyloxy-phenyl)-piperazine

[0858]

[0859] The title compound was prepared in a manner similar toPreparation 32A except that 3-methylphenyl boronic acid was used. LRMS(ESI+) 269 (M+1)

Preparation 34A 1-(2-p-tolyloxy-phenyl)-piperazine

[0860]

[0861] The title compound was prepared in a manner similar toPreparation 32A except that 4-methylphenylboronic acid was used. LRMS(ESI+) 269 (M+1)

Preparation 35A 1-[2-(3-chloro-phenoxy)-phenyl]-piperazine

[0862]

[0863] The title compound was prepared in a manner similar toPreparation 32A except that 3-chlorophenylboronic acid was used. LRMS(ESI+) 289 (M+1)

Preparation 36A 1-[2-(3-methoxy-phenoxy)-phenyl]-piperazine

[0864]

[0865] The title compound was prepared in a manner similar toPreparation 32A except that 3-methoxyphenylboronic acid was used. LRMS(ESI+) 285 (M+1)

Preparation 37A (Benzylamine from Nitrile Reduction)1-Boc-4-(2-aminomethyl-phenyl)-piperazine

[0866]

[0867] To a solution of (2-cyano-phenyl)-piperazine (2.4 g, 12.78 mmol)in THF and H₂O (25 mL, 1:1) was added K₂CO₃ (3.9 g, 28.12 mmol). Thesolution was allowed to stir for about 10 minutes at r.t. Boc-anhydride(3.1 g, 14.06 mmol) was then added and reaction was allowed to stir for1 h. The reaction mixture was diluted with EtOAc (100 mL) and washedwith sat. NaHCO₃ (100 mL) and brine (100 mL). The organic phase wasconcentrated to dryness yielding 3.2 g of1-Boc-4-(2-cyano-phenyl)-piperazine (88%). To a solution of sodiumborohydride (2.1 g, 56.03 mmol) in THF (25 mL) at 0° C. was added TFA(4.3 mL, 56.03 mmol) dropwise. 1-Boc-4-(2-cyano-phenyl)-piperazine (3.2g, 11.21 mmol) was then added slowly at r.t. The reaction was allowed tostir for about 12 hours at r.t. The reaction was quenched with H₂O,diluted five-fold with EtOAc and washed with brine. The organic phasewas concentrated to dryness yielding about 1.0 g of1-Boc-4-(2-aminomethyl-phenyl)-piperazine (30%). MS (ESI+) 292.1 (M+1)

Preparation 38A 1-Boc-4-(2-dimethylaminomethyl-phenyl)-piperazine

[0868]

[0869] 1-Boc-4-(2-aminomethyl-phenyl)-piperazine (2.0 g, 6.86 mmol) wasdissolved in CH₃CN (15 mL) and cooled to about 0° C. Aqueousformaldehyde (37% wt. in H₂O) (7.56 mL) was added to the cold solutionfollowed by the addition of sodium cyanoborohydride (2.15 g, 34.32mmol). The reaction mixture was allowed to stir at 0° C. for about 5minutes and then allowed to naturally warm to room temperature. Themixture was then concentrated to dryness. The resulting residue wastaken up in EtOAc (100 mL) and washed with saturated NaHCO₃ solution(100 mL) and brine (100 mL). The organic phase was concentrated todryness to afford about 2.2 g of crude material. MS (ESI+) 320.2 [M+1]

Preparation 39A1-Boc-4-[2-(methanesulfonylamino-methyl)-phenyl]-piperazine

[0870]

[0871] 1-Boc-4-(2-aminomethyl-phenyl)-piperazine (2.09 g, 7.18 mmol) wasdissolved in methylene chloride (50 mL), cooled to 0° C. and treatedwith triethylamine (1.5 mL, 10.8 mmol) followed by methanesulfonylchloride (0.67 mL, 8.61 mmol). The resulting mixture was stirred forabout 3 hours at r.t., and then diluted with ether (200 mL) and washedwith water (50 mL), saturated aqueous sodium bicarbonate (50 mL) andbrine (50 mL), which is then dried over anhydrous magnesium sulfate.Concentration under reduced pressure followed by silica gelchromatography (30% ethyl acetate in hexanes) afforded the titlecompound (2.07 g, 78%) as a clear oil.

[0872]¹H NMR (CDCl₃) δ7.25-7.40 (m, 2H), 7.00-7.15 (m, 2H), 4.40 (s,1H), 3.55-3.65 (m, 4H), 2.80-2.95 (m, 4H), 2.75 (s, 3H), 1.60 (s, 9H).TLC (SiO₂): R_(f)=0.50 (50% EtOAc/hexanes)

Preparation 40A 1-Boc-4-[2-(acetylamino-methyl)-phenyl]-piperazine

[0873]

[0874] The title compound was Prepared in a similar manner toPreparation 39A except that acetic anhydride was used instead ofmethanesulfonyl chloride.

[0875]¹H NMR (CDCl₃) δ7.45-7.55 (m, 2H), 7.05-7.15 (m, 2H), 6.20 (s,1H), 4.45-4.50 (m, 2H), 3.55-3.65 (m, 4H), 2.75-2.90 (m, 4H), 2.05 (s,3H), 1.60 (s, 9H). TLC (SiO₂): R_(f)=0.15 (50% EtOAc/hexanes)

Preparation 41A1-Boc-4-[2-(bezenesulfonylamino-methyl)-phenyl]-piperazine

[0876]

[0877] The title compound was Prepared in a similar manner toPreparation 39A except that benzenesulfonyl chloride was used instead ofmethanesulfonyl chloride.

[0878]¹H NMR (CDCl₃) δ6.90-7.90 (m, 9H), 5.75-5.85 (m, 1H), 4.15-4.25(m, 2H), 3.50-3.60 (m, 4H), 2.60-2.75 (m, 4H), 1.20-1.55 (m, 9H). TLC(SiO₂): R_(f)=0.85 (100% EtOAc)

Preparation 42A1-Boc-4-[2-(ethanesulfonylamino-methyl)-phenyl]-piperazine

[0879]

[0880] The title compound as Prepared in a similar manner to Preparation39A except that ethanesulfonyl chloride was used instead ofmethanesulfonyl chloride.

[0881]¹H NMR (CDCl₃) δ7.05-7.35 (m, 4H), 4.35-4.45 (m, 2H), 3.70-3.80(m, 5H) 2.85-2.90 (m, 6H), 1.25-1.50 (m, 12H). TLC (SiO₂): R_(f)=0.85(100% EtOAc)

Preparation 43A1-Boc-4-[2-(propane-2-sulfonylamino-methyl)-phenyl]-piperazine

[0882]

[0883] The title compound was Prepared in a similar manner toPreparation 39A except that isopropylsulfonyl chloride was used insteadof methanesulfonyl chloride.

[0884]¹H NMR (CDCl₃) δ7.00-7.35 (m, 4H), 4.45-4.50 (m, 1H), 3.75-3.85(m, 4H), 2.90-3.00 (m, 4H), 1.95-2.25 (m, 8H), 1.20-1.55 (m, 10H)

Preparation 44A 1-Boc-4-[2-(isobutyrylamino-methyl)-phenyl]-piperazine

[0885]

[0886] The title compound was Prepared in a similar manner toPreparation 39A except isobutyryl chloride was used instead ofmethanesulfonyl chloride and diisopropylethylamine was used as the base.

[0887]¹H NMR (CDCl₃) δ7.34-7.41 (m, 2H) 7.14-7.22 (m, 2 H), 6.39-6.47(m, 1 H), 4.53-4.58 (m, 2 H), 2.78-2.95 (m, 4 H), 2.76-2.87 (m, 4 H),1.43-1.54 (s, 9H) 1.15-1.21 (m, 6H)

Preparation 45A [2-(propionylamino-methyl)-phenyl]-piperazine

[0888]

[0889] 1-Boc-4-(2-aminomethyl-phenyl)-piperazine (0.75 g, 2.6 mmol) wasdissolved in methylene chloride (20 mL), treated with DIPEA (2.3 mL, 13mmol), and cooled to about 0° C. Propionyl chloride (0.20 mL, 2.34 mmol)was added and the mixture was stirred for about 1 hour at 0° C. andsubsequently stirred overnight at r.t. The mixture is diluted with ethylacetate (400 mL), washed with water (45 mL), saturated aqueous sodiumbicarbonate (45 mL) and brine (45 mL), and then dried over anhydroussodium sulfate. Concentration under reduced pressure followed by silicagel chromatography (50% ethyl acetate in hexanes) gave an oil, which wasdissolved in methylene chloride (10 mL). The mixture was stirred withTFA (10 mL) for about 1.5 hours. The mixture was concentrated underreduced pressure, and the residue taken up in water (25 mL). Sodiumhydroxide (1.0 g, 25 mmol) and ethyl acetate (25 mL) were added and themixture was stirred for about 45 minutes. The organic phase wascollected and the aqueous phase was extracted with ethyl acetate (45mL). The combined organic fractions were washed with water (20 mL) andbrine (20 mL) and then dried over anhydrous sodium sulfate. The solventwas concentrated under reduced pressure to yield the title compound(0.26 g, 40%) as a clear oil.

[0890]¹H NMR (CDCl₃) δ6.99-7.43 (m, 5 H), 6.46-6.71 (bs, 1 H), 4.46-4.72(s, 2 H), 2.79-3.23 (m, 8 H), 2.14-2.43 (m, 2 H), 1.07-1.38 (m, 3 H).

[0891] Alternatively, the title compound was prepared in the followingprocedure: About 0.40 g (1.37 mmol) of1-Boc-4-(2-aminomethyl-phenyl)-piperazine, 0.11 ml of (1.51 mmol)propionic acid, 0.22 g (1.64 mmol) of HOBt, 0.31 g (1.64 mmol) of EDC,and 0.24 ml (1.37 mmol) of DIEA were mixed in 30 ml THF under nitrogenand stirred overnight at r.t. The reaction was concentrated to drynessand ethyl acetate was added. The mixture was washed with saturatedbicarbonate and brine, and then dried with sodium sulfate. The residuewas purified by flash chromatography eluting with 1:1 hexane/ethylacetate giving about 0.4 g (86% yield). The material was deprotectedusing TFA/DCM to give 4-[2-(propionylamino-methyl)-phenyl]-piperazine.LRMS (ESI+): 248 (M+1)

Preparation 46A4-{2-[(2,2-Dimethyl-propionylamino)-methyl]-phenyl}-piperazine

[0892]

[0893] 1-Boc-4-(2-Aminomethyl-phenyl)-piperazine (0.75 g, 2.6 mmol) wasdissolved in methylene chloride (20 mL). DIPEA (2.3 mL, 13 mmol) wasadded, and the mixture was cooled to about 0° C. The solution wastreated with trimethylacetyl chloride (0.28 g, 0.28 mL, 2.3 mmol) andstirred for about 1 hour at 0° C. The solution was warmed to r.t. andstirred overnight. The mixture was diluted with ethyl acetate (400 mL),washed with water (60 mL), saturated aqueous sodium bicarbonate (60 mL)and brine (60 mL) and then dried over anhydrous sodium sulfate. Thesolution was concentrated under reduced pressure and purified via silicagel chromatography (80% ethyl acetate in hexanes) to afford a clear oil,which was subsequently stirred in neat TFA (5 mL) for about 1 hours. Thesolvent was evaporated under reduced pressure and the residue taken upin water (30 mL). Sodium hydroxide (1 g, 25 mmol) and ethyl acetate (30mL) were added, and the mixture was stirred for about 45 minutes. Theorganic phase was collected and the aqueous phase was extracted withethyl acetate (60 mL). The combined organic fractions were washed withwater (45 mL) and brine (30 mL) and then dried over anhydrous sodiumsulfate. Concentration under reduced pressure afforded the titlecompound (0.54 g, 75%) as a clear oil.

[0894]¹H NMR (CDCl₃) δ6.97-7.36 (m, 4H), 6.63-6.86 (bs, 1H), 4.47-4.65(m, 2H), 2.66-3.24 (m, 8H), 1.18 (s, 9H).

Preparation 47A 4-[2-(benzoylamino-methyl)-phenyl]-piperazine

[0895]

[0896] 1-Boc-4-(2-Aminomethyl-phenyl)-piperazine (0.47 g, 1.6 mmol) wasdissolved in methylene chloride (20 mL). DIPEA (1.5 mL, 8.5 mmol) wasadded, and the mixture was cooled to about 0° C. The mixture was treatedwith benzoyl chloride (0.20 g, 0.16 mL, 1.4 mmol). The resulting mixturewas stirred for about 1 hour at 0° C., and then warmed to r.t. andstirred overnight. The mixture was diluted with ethyl acetate (500 mL),washed with water (45 mL), saturated aqueous sodium bicarbonate (45 mL)and brine (45 mL) and then dried over anhydrous sodium sulfate. Thesolution was concentrated under reduced pressure and purified via silicagel chromatography (50% ethyl acetate in hexanes) to afford a clear oil,which was subsequently stirred in neat TFA (5 mL) for about 1 hour. Thesolvent was evaporated under reduced pressure to afford the titlecompound as a clear oil (0.30 g, 100%).

[0897]¹H NMR (CDCl₃) δ7.32-7.84 (m, 9H), 4.71-4.86 (m, 2H), 3.42-3.65(m, 4H), 3.24-3.42 (m, 4H).

Preparation 48A1-Boc-4-{2-[(methanesulfonyl-methyl-amino)-methyl]-phenyl}-piperazine

[0898]

[0899] To a stirred suspension of sodium hydride (60% in oil, 113 mg,2.82 mmol) in THF (20 mL) at 0° C. under nitrogen was added a solutionof 1-Boc-4-[2-(methanesulfonylamino-methyl)-phenyl]-piperazine (0.99 g,2.68 mmol) in THF (5 mL). The mixture was stirred for about 1 hour atroom temperature. It then was cooled back to 0° C. and treated withmethyl iodide (0.184 mL, 2.95 mmol). After stirring for about 20 hours,the reaction mixture was diluted with ether (150 mL) and then quenchedby addition of saturated aqueous ammonium chloride (50 mL). The organicphase was separated, washed with water (50 mL) and brine (50 mL) andthen dried over magnesium sulfate. Concentration under reduced pressurefollowed by silica gel chromatography (30% ethyl acetate in hexanes)afforded the title compound (0.96 g, 94%) as a clear oil.

[0900]¹H NMR (CDCl₃) δ7.45-7.55 (m, 1H), 6.95-7.35 (m, 3H), 4.45 (s,2H), 3.45-3.60 (m, 4H), 3.05 (s, 3H), 2.75-2.90 (m, 4H), 2.75 (s, 3H),1.60 (s, 9H). TLC (SiO₂): R_(f)=0.70 (50% EtOAc/hexanes).

Preparation 49A1-Boc-4-{2-[(benzyl-methanesulfonyl-amino)-methyl]-phenyl}-piperazine

[0901]

[0902] The title compound was prepared in a similar manner toPreparation 48A except that benzyl bromide was used.

[0903]¹H NMR (CDCl₃) δ7.70-7.75 (m, 1H), 7.25-7.55 (m, 8H), 4.75 (s,2H), 4.50 (s, 2H), 3.45-3.60 (m, 4H), 3.05 (s, 3H), 2.75-2.90 (m, 4H),1.65 (s, 9H). TLC (SiO₂): R_(f)=0.70 (50% EtOAc/hexanes).

Preparation 50A1-Boc-4-{2-[(ethyl-methanesulfonyl-amino)-methyl]-phenyl}-piperazine

[0904]

[0905] The title compound was prepared in a similar manner toPreparation 48A except that ethyl iodide was used. TLC (SiO₂):R_(f)=0.25 (30% EtOAc/hexanes).

Preparation 51A1-Boc-4-{2-[(Acetyl-methyl-amino)-methyl]-phenyl}-piperazine

[0906]

[0907] The title compound was prepared in a similar manner toPreparation 48A starting with1-Boc-4-[2-(acetylamino-methyl)-phenyl]-piperazine (0.58 g, 1.7 mmol).The title compound was obtained (0.36 g, 60%) as a clear oil. TLC(SiO₂): R_(f)=0.33 (66% ethyl acetate in hexanes).

Preparation 52A1-Boc-4-{2-[(Acetyl-benzyl-amino)-methyl]-phenyl}-piperazine

[0908]

[0909] The title compound was prepared in a similar manner toPreparation 51A except that benzyl bromide was used. TLC (SiO₂):R_(f)=0.20 (66% ethyl acetate in hexanes)

Preparation 53A1-Boc-4-{2-[(Acetyl-ethyl-amino)-methyl]-phenyl}-piperazine

[0910]

[0911] The title compound was prepared in a similar manner toPreparation 51A except that ethyl iodide was used. TLC (SiO₂):R_(f)=0.35 (66% ethyl acetate in hexanes).

Preparation 54A (Benzylamine from Benzylic Alcohol Via Mitsunobu)1-Boc-4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazine

[0912]

[0913] Step 1: 1-Boc-4-(2-carboxy-phenyl)-piperazine

[0914] To a solution of 1-(2-cyanophenyl)-piperazine (7.5 g, 40 mmol) in100 mL of absolute ethanol was added 200 mL of 25% aqueous KOH. Thesolution was heated to reflux for about 48 hours and then cooled toabout 0° C. The solution was acidified with 180 mL of 5 M HCl and thensolid NaHCO₃ was added to bring the pH of the solution to about 10.After concentration in vacuo to remove 60 mL of solvent, dioxane (300mL), NaHCO₃ (12.7 g, 120 mmol) and Boc₂O (11.4 g, 52.2 mmol) were added.The solution was stirred overnight and then acidified with 5 M HCl toabout pH 1. After separation, the aqueous solution was extracted withEtOAc (3×). The combined organic solutions were washed with water (2×),brine and then dried (Na₂SO₄), filtered and concentrated to give thetitle compound. LRMS (ES−): 305.2 (M−1)

[0915] Step 2: 1-Boc-4-(2-hydroxymethyl-phenyl)-piperazine

[0916] To a solution of 1-Boc-4-(2-carboxy-phenyl)-piperazine from Step1 in 340 mL of THF at 0° C. was added BH₃-THF (120 mL of a 1 M solutionin THF). The cold bath was removed, and the solution stirred overnight.The solution was cooled to about 0° C. and then 60 mL of 2 M NaOH wasadded followed by EtOAc and brine. After separation, the aqueoussolution was extracted with EtOAc (3×). The combined organic solutionswere washed with water (2×) and brine, and then dried (Na₂SO₄), filteredand concentrated to give about 11.2 g (38.3 mmol, 96%) of the titlecompound. LRMS (ESI+): 393.2 [M+1]

[0917] Step 3: To a solution of1-Boc-4-(hydroxymethyl-phenyl)-piperazine (300 mg, 1.02 mmol, 1.0 eq.),1,2,4 triazole (104 mg, 1.53 mmol, 1.5 eq.), triphenylphosphine (535 mg,2.04 mmol, 2.0 eq.) and THF at 0° C. under nitrogen was added DEAD(0.321 mL, 2.04 mmol, 2.0 eq.) slowly so that temperature of reactiondoes not rise above 10° C. After addition was completed, the ice bathwas removed and the reaction mixture was stirred at r.t. overnight.Methanol was added and the mixture was stirred for about 15 minutes. Themixture was then concentrated. Purification by flash chromatography (35g SiO₂, linear gradient 50-70% EtOAc/Hexane for 15 minutes and 70% EtOAcfor 18 minutes) afforded Boc protected title compound (200 mg, 0.5 mmol,57%). LRMS (ESI+): 344.1 (M+1)

Preparation 55A 1-Boc-4-(2-tetrazol-2-ylmethyl-phenyl)-piperazine

[0918]

[0919] The title compound was prepared in a manner similar toPreparation 54A except that tetrazole was used. LRMS (ESI+): 289.1(M-Boc).

Preparation 56A 1-Boc-4-(2-imidazol-1-ylmethyl-phenyl)-piperazine

[0920]

[0921] The title compound was prepared in a manner similar toPreparation 54A except that imidazole was used. LRMS (ESI+): 343.2 (M+1)

Preparation 57A 1-Boc-4-(2-azidomethyl-phenyl)-piperazine

[0922]

[0923] 1-Boc-4-(2-Hydroxymethyl-phenyl)-piperazine (4.59 g, 15.7 mmol)was dissolved in toluene (75 mL). Triphenylphosphine (8.3 g, 31.6 mmol)was added followed by zinc azide pyridine salt (3.61 g, 11.72 mmol).Diisopropyl azodicarboxylate (6.27 mL, 31.6 mmol) was added dropwise,and the solution was stirred at r.t. for about 12 hours. The mixture wasconcentrated under reduced pressure and purified using silicachromatography (12% ethyl acetate in hexanes) to give the title compound(1.89 g, 51%) as an oil.

[0924]¹H NMR (CDCl₃) δ7.35-7.05 (m, 4H), 4.45 (s, 2H), 3.60-3.50 (m,4H), 2.85-2.75 (m, 4H), 1.50 (s, 9H).

Preparation 58A1-Boc-4-[2-(4-Methoxycarbonyl-[1,2,3]triazol-1-ylmethyl)-phenyl]-piperazine

[0925]

[0926] 1-Boc-4-(2-Azidomethyl-phenyl)-piperazine (0.25 g, 0.79 mmol) wasdissolved in deuterated chloroform (3 mL). Methyl propiolate (0.35 mL,3.9 mmol) was added, and the mixture was heated to reflux for about 4hours and then cooled to r.t. The mixture was concentrated under reducedpressure and purified using silica chromatography (50% ethyl acetate inhexanes) to give the title compound (0.155 g, 49%) as an oil.

[0927]¹H NMR (CDCl₃) δ7.35-7.05 (m, 4H), 5.75 (s, 2H), 3.95 (s, 3H),3.55-3.45 (m, 4H), 2.80-2.70 (m, 4H), 3.80-3.85 (m, 1H), 1.50 (s, 9H).

Preparation 59A1-Boc-4-[2-(4-tert-Butyl-[1,2,3]triazol-1-ylmethyl)-phenyl]-piperazine

[0928]

[0929] In a sealed tube,4-(2-Azidomethyl-phenyl)-piperazine-1-carboxylic acid tert-butyl ester(0.366 g, 1.15 mmol) was dissolved in toluene (5 mL).3,3-dimethyl-1-butyne (0.7 mL, 5.64 mmol) was added and the mixture washeated to reflux for about 48 hours and then cooled to r.t. The mixturewas concentrated under reduced pressure and purified using silicachromatography (50% ethyl acetate in hexanes) to give the title compound(0.212 g, 60%) as an oil.

[0930]¹H NMR (CDCl₃) δ7.35-7.05 (m, 4H), 5.75 (s, 2H), 3.60-3.45 (m,4H), 2.80-2.70 (m, 4H), 1.50 (s, 9H), 1.35 (s, 9H).

Preparation 60A (Benzylamine from Benzylic Alcohol Via Mesylate)1-Boc-4-[2-(3R-dimethylamino-pyrrolidin-1-ylmethyl)-phenyl-piperazine

[0931]

[0932] To a solution of 1-Boc-4-(2-hydroxymethyl-phenyl)piperazine (300mg, 1.03 mmol, 1.0 eq.), triethylamine (0.17 mL, 1.2 mmol, 1.2 eq.),DMAP (6 mg, 0.05 mmol, 0.05 eq.) in CH₂Cl₂ (10 mL) was addedmethanesulfonyl chloride (0.085 mL, 1.1 mmol, 1.1 eq.). The solution wasstirred at r.t. under N₂ for about 2 hours. A solution of3R-3-(dimethylamino)pyrrolidine (0.63 mL, 5.0 mmol, 5.0 eq.) in THF (3mL) was added, and the mixture was allowed to stir at r.t. overnight.The mixture was diluted with CH₂C₂ (10 mL) and washed with saturatedaqueous NaHCO₃ (15 mL) and brine (15 mL). The aqueous layers wereextracted with CH₂Cl₂ (3×). The combined organic extracts were dried(Na₂SO₄), filtered and concentrated. Purification by flashchromatography (35 g SiO₂, 40 ml/min, linear gradient 0-10% 2.0 M NH₃ inMeOH/CH₂Cl₂ for 25 minutes and 10% 2.0M NH₃ in MeOH/CH₂Cl₂ for 7minutes) afforded the title compound as a white solid (280 mg, 0.72mmol, 72%). LRMS (ESI+): 389.2 [M+1]

Preparation 61A1-Boc-4-[2-(3S-dimethylamino-pyrrolidin-1-ylmethyl)-phenyl-piperazine

[0933]

[0934] The title compound was prepared in a manner similar toPreparation 60A except that 3S-3-(dimethylamino)pyrrolidine was used.LRMS (ESI+): 389.2 (M+1)

Preparation 62A 1-Boc-4-(2-pyrrolidin-1-ylmethyl-phenyl)-piperazine

[0935]

[0936] The title compound was prepared in a manner similar toPreparation 60A except that pyrrolidine was used. LRMS (ESI+): 246.1(M+1)

Preparation 63A1-Boc-4-[2-(2-methyl-imidazol-1-ylmethyl)-phenyl]-piperazine

[0937]

[0938] The title compound was prepared in a manner similar toPreparation 60A except that 2-methyl-imidazole was used. LRMS (ESI+):357.2 (M+1)

Preparation 64A1-Boc-4-[2-(2-ethyl-imidazol-1-ylmethyl)-phenyl]-piperazine

[0939]

[0940] The title compound was prepared in a manner similar toPreparation 60A except that 2-isopropyl-imidazole was used. LRMS (ESI+):371.3 (M+1)

Preparation 65A1-Boc-4-[2-(2-ethyl-imidazol-1-ylmethyl)-phenyl]-piperazine

[0941]

[0942] The title compound was prepared in a manner similar toPreparation 60A except that 2-ethyl-imidazole was used. LRMS (ESI+):385.2 (M+1)

Preparation 66A1-Boc-4-[2-(2-methylsulfanyl-imidazol-1-ylmethyl)-phenyl]-piperazine

[0943]

[0944] The title compound was prepared in a manner similar toPreparation 60A except that 2-ethylsulfanyl-1H-imidazole was used. LRMS(ESI+): 403.3 (M+1)

Preparation 67A1-Boc-4-(5-methyl-2-pyrrolidin-1-ylmethyl-phenyl)-piperazine

[0945]

[0946] To a solution of 2-bromo-4-methylaniline (558 mg, 3.0 mmol) in 30mL of acetbnitrile was added tetrafluoroborate (600 μL of a 54% solutionin Et₂O, 4.35 mmol). The solution was cooled to about 0° C. and t-butylnitrite (55 uL, 4.62 mmol) was added. After stirring for about 45minutes, the solution was transferred to a solution of CuCN (800 mg,8.93 mmol) and NaCN (1.47 g, 30 mmol) in 30 mL, of water cooled to 0° C.via cannula. The cold bath was removed. After stirring overnight, theaqueous solution was extracted with Et₂O (2×). The combined organicsolutions were washed with 1 M HCl, saturated sodium bicarbonate, waterand brine, and then dried (Na₂SO₄), filtered and concentrated. Thematerial was adsorbed onto 3 g of silica gel and purified by silica gelflash chromatography (4×15 cm column, 5-20 Et₂O/pentane, over 48 min at35 mL/min) to afford about 320 mg (1.63 mmol, 54%) of2-bromo-4-methyl-benzonitrile as a colorless oil. GC/MS (EI): 195.

[0947] 2-Bromo-4-methyl-benzonitrile was coupled to piperazine usingPreparation 1A Buchwald chemistry to afford4-(2-cyano-5-methyl-phenyl)-piperazine.4-(2-cyano-5-methyl-phenyl)-piperazine is converted to4(2-hydroxymethyl-5-methyl-phenyl)-piperazine in a manner similar topreparation 54A Steps 1 and2.4-(2-Hydroxymethyl-5-methyl-phenyl)-piperazine was converted to thetitle compound in a manner similar to Preparation 60A except thatpyrrolidine was used to displace the mesylate. LRMS (ESI+): 360.3 (M+1)

Preparation 68A1-Boc-4-(5-isopropyl-2-pyrrolidin-1-ylmethyl-phenyl)-piperazine

[0948]

[0949] The title compound was prepared in a manner similar to thePreparation 67A except that 2-bromo-4-isopropylaniline was used as thestarting material. LRMS (ESI+): 388.3 (M+1)

Preparation 69A1-Boc-4-(2-dimethylaminomethyl-5-trifluoromethyl-phenyl)piperazine

[0950]

[0951] To a solution of piperazine (13.7 g, 159 mmol) in 20 mL DMSO wasadded 2-fluoro-4-trifluoromethylbenzonitrile (10 g, 52.9 mmol). Afterstirring overnight, the solution was diluted with 200 mL of EtOAc,washed with water and brine, and then dried (Na₂SO₄), filtered andconcentrated to afford about 13.0 g (51.1 mmol, 96%) of4-(2-cyano-5-trifluoromethyl-phenyl)piperazine. LRMS (ESI+): 256.1[M+1]. The tide compound was prepared from4-(2-cyano-5-trifluoromethyl-phenyl)piperazine in the same manner asdescribed in Preparation 67A except that dimethyl amine was used todisplace the mesylate. LRMS (ESI+): 388.1 [M+1]

Preparation 70A1-Boc-4-(2-pyrrolidin-1-ylmethyl-5-trifluoromethyl-phenyl)-piperazine

[0952]

[0953] The title compound was synthesized in similar manner as describedin Preparation 69A except that pyrrolidine was used to displace themesylate. LRMS (ESI+): 414.3 (M+1)

Preparation 71A1-Boc-4-(2-pyrrolidin-1-ylmethyl-4-trifluoromethyl-phenyl)-piperazine

[0954]

[0955] The title compound was synthesized in similar manner as describedin Preparation 70A except that 2-fluoro-5-trifluoromethylbenzonitrilewas used as the starting material. LRMS (ESI+): 414.3 (M+1)

Preparation 72A1-Boc-4-(2-pyirrolidin-1-ylmethyl-6-trifluoromethyl-phenyl)-piperazine

[0956]

[0957] The title compound was synthesized in similar manner as describedin Preparation 70A except that 2-fluoro-3-trifluoromethylbenzonitrilewas used as the starting material. LRMS (ESI+): 414.3 (M+1)

Preparation 73A1-Boc-4-(2-pyrrolidin-1-ylmethyl-3-trifluoromethyl-phenyl)-piperazine

[0958]

[0959] 4-(2-cyano-3-trifluoromethyl-phenyl)piperazine was prepared in amanner similar to 4-(2-cyano-5-trifluoromethyl-phenyl)piperazinedescribed above except that 2-fluoro-6-trifluoromethylbenzonitrile wasused as the starting material. To a solution of4-(2-cyano-3-trifluoromethyl-phenyl)piperazine (1.35 g, 5.29 mmol, 1.0eq.) in dioxane (40 mL) was added a solution of DIBAL in heptane (1.0 Min heptane, 13.2 mL, 13.22 mmol, 2.5 eq.). The resulting mixture wasstirred at r.t. for about 3 days. The mixture was transferred viacannula to 0.5 M Rochelle salt and stirred for about 2 hours. NaHCO₃(1.3 g, 15.9 mmol, 3.0 eq) and di-tert-butyl dicarbonate (1.7 g, 7.29mmol, 1.5 eq.) were added and the mixture was stirred at r.t. overnight.The mixture was partitioned between EtOAc (100 mL) and brine (50 mL).The organic layer was separated, and aqueous layer was extracted withEtOAc (2×). The combined organic extracts were washed with H₂O andbrine, and then dried (Na₂SO₄), filtered and concentrated. Purificationby flash chromatography (120 g SiO₂, 40 mL/min, linear gradient 0-25%EtOAc/Hexane for 10 minutes and 25% EtOAc/Hexane for 23 minutes)afforded N-boc-4-(2-formyl-3-trifluoromethyl-phenyl)piperazine (637 mg,1.77 mmol, 35%). LRMS (ESI+): 359.1 [M+1]

[0960] To a solution ofN-boc-4-(2-formyl-3-trifluoromethyl-phenyl)piperazine (358 mg, 1 mmol,1.0 eq.) in MeOH (10 mL) was added pyrrolidine (0.093 mL, 1.1 mmol, 1.1eq.). The mixture was refluxed overnight. The reaction was cooled toabout 0° C., and NaBH₄ on alumina (10 wt % on basic alumina, 570 mg, 1.5mmol, 1.5 eq.) was added. After the addition was compete, the ice bathwas removed, and the mixture was stirred at r.t. for about 2 hours. Themixture was filtered through celite, washed with methanol andconcentrated. The solution was diluted with EtOAc (50 mL) and washedwith saturated NaHCO₃ and brine. The aqueous layers were extracted withEtOAc (2×). The combined organic extracts were dried (Na₂SO₄), filteredand concentrated. Purification by flash chromatography (35 g SiO₂, 40mL/min, linear gradient 0-10% MeOH/CH₂Cl₂ for 25 minutes and 10%MeOH/CH₂Cl₂ for 7 minutes) gave the title compound (298 mg, 0.72 mmol,72%). LRMS (ESI+): 414.3 (M+1)

Preparation 74A (Derivatives of 1-Boc-4-(2-amino-phenyl)-piperazine)1-Boc-4-(2-amino-phenyl)-piperazine

[0961]

[0962] To a solution of N-(2-nitrophenyl)-piperazine (30 g, 145 mmol)and triethylamine (28.3 mL, 203 mmol) in 600 mL of CH₂Cl₂ was addedBoc₂O (38 g, 174 mmol). After stirring overnight, the solution waswashed with saturated aqueous sodium bicarbonate and brine, and thendried (Na₂SO₄), filtered and concentrated to afford an orange oil. To asolution of the oil in 2 L of ethanol was added 6 g of 5% Pd/C. Aftershaking under 60 psi H₂ overnight, the solution was filtered andconcentrated to afford about 39 g (140 mmol, 97%) of1-boc-4-(2-aminophenyl)-piperazine as abrown solid. LRMS: 278.1 (M+1)

Preparation 75A 1-Boc-4-(2-dimethylaminophenyl)piperazine

[0963]

[0964] To a solution of 1-boc-4-(2-nitrophenyl) piperazine (500 mg, 1.63mmol, 1.0 eq) in IPA (20 mL) was added formaldehyde (3.3 mL 37% solutionin H₂O, 4.07 mmol, 2.5 eq) and 10% Pd/C (125 mg, 25 wt %). The mixturewas shaken under hydrogen at 60-psi overnight. The mixture was filteredand diluted with CH₂Cl₂. The aqueous solution was separated, and theorganic solution was dried (Na₂SO₄), filtered through a pad of celiteand concentrated. Purification by flash chromatography (35 g SiO₂, 40ml/min, linear gradient 0-15% EtOAc/hexane for 20 minutes and 15%EtOAc/hexane for 13 minutes) gave about 480 mg (1.57 mmol, 97%) of thetitle compound as a solid. LRMS (ESI+): 306.2 (M+1)

Preparation 76A 1-Boc-4-[2-(isobutylamido)-phenyl]-piperazine

[0965]

[0966] To a solution of 1-Boc-4-(2-amino-phenyl)-piperazine (2.77 g, 10mmol), triethyl amine (2.8 mL, 20 mmol), and DMAP (70 mg, 0.57 mmol) in50 mL of CH₂Cl₂ was added isobutyryl chloride (1.15 mL, 11 mmol). Afterstirring overnight, saturated aqueous sodium bicarbonate was added andthe solution was concentrated. The solution was diluted with EtOAc,washed with 1 M HCl, water, saturated aqueous sodium bicarbonate andbrine, and then dried (Na₂SO₄), filtered and concentrated to affordabout 3.29 g (9.4 mmol, 94%) of the title compound. LRMS: 348.2 (M+1)

Preparation 77A 1-Boc-4-[2-(3-methyl-butyrylamino)-phenyl]-piperazine

[0967]

[0968] The title compound was prepared in a manner similar toPreparation 76A except that isovaleryl chloride was used instead ofisobutyryl chloride. LRMS (ESI+): 362.2 (M+1)

Preparation 78A 1-Boc-4-(2-isobutylamino-phenyl)-piperazine

[0969]

[0970] To a solution of 1-Boc-4-(2-isobutylamino-phenyl)-piperazine(2.72 g, 7.8 mmol) in 50 mL of THF was added BH₃-THF (24 mL of 1 Msolution in THF, 24 mmol). After stirring for about 1 hour at 60° C.,the solution was cooled to r.t. and then 25 mL of 1 M NaOH was added.After stirring for about 2 hours, brine and EtOAc were added. Theorganic solution was washed with water (2×) and brine, and then dried(Na₂SO₄), filtered and concentrated. Purification by flashchromatography (Biotage 40L column, 0 to 30% EtOAc/Hex linear gradientover 48 min at 35 mL/min) afforded about 2.35 g (7.05 mmol, 90%) of thetitle compound. LRMS: 334.2 (M+1)

Preparation 79A 1-Boc-4-(2-methanesulfonylamino-phenyl)-piperazine

[0971]

[0972] To a solution of 1-boc-4-(2-aminophenyl)-piperazine (5.55 g, 20mmol) and triethyamine (5.6 mL, 40 mmol) in 200 mL of CH₂Cl₂ was addedmethanesulfonyl chloride (1.55 mL, 20 mmol). After stirring for about 4hours, the solution was concentrated, and the residue dissolved in 200mL of EtOAc. The solution was washed with 1 M HCl (2×), water and brine,and then dried (Na₂SO₄), filtered and concentrated to afford about 6.68g (18.8 mmol, 94%) of the title compound as a brown solid. LRMS: 356.1(M+1)

Preparation 80A 1-Boc-4-[2-(3,3-Dimethyl-ureido)-phenyl]-piperazine

[0973]

[0974] To a solution of 1-Boc-4-(2-aminophenyl)-piperazine (270 mg, 1.0mmol) and Et₃N (400 microliter, 2.89 mmol) in 10 mL of CH₂Cl₂ was addeddimethylcarbamyl chloride (135 microliter, 1.48 mmol). After stirringfor about 1 hour, DMAP (10 mg) was added. After stirring for about 3days, another 800 microliter of Et₃N and 270 microliter ofdimethylcarbamyl chloride were added. After stirring overnight, thesolution was diluted with EtOAc, washed with 1 M HCl (2×), saturatedsodium bicarbonate, water and brine, and then dried (Na₂SO₄), filteredand concentrated. Purification by silica gel chromatography (35 g SiO₂,20 to 50% EtOAc/hexanes, over 30 minutes at 35 mL/min) afforded about 20mg (0.057 mmol, 6%) of the tide compound as a white solid. LRMS: 349.2(M+1)

Preparation 81A 1-Boc-4-[2-(3-isopropyl-ureido)-phenyl]-piperazine

[0975]

[0976] To a solution of 1-Boc-4-(2-aminophenyl)-piperazine (270 mg, 1.0mmol) in 10 mL of THF was added isopropyl isocyanate (90 uL, 1.46 mmol).After stirring for about 1 hour, another 90 microliter of isopropylisocyanate was added. After stirring for about 3 days, another 290microliter of isopropyl isocyanate was added. After stirring overnight,the solution was concentrated. Purification by silica gel chromatography(35 g SiO₂, 20 to 50% EtOAc/hexanes, over 30 minutes at 35 mL/min)afforded about 240 mg (0.66 mmol, 66%) of the title compound as a whitesolid. LRMS: 363.2 (M+1)

Preparation 82A1-Boc-4-[2-(isobutyl-methanesulfonyl-amino)-phenyl]-piperazine

[0977]

[0978] To a solution of1-boc-4-(2-methanesulfonylamino-phenyl)-piperazine (1.07 g, 3.0 mmol) in50 mL of DMF was added NaH (240 mg of a 60% dispersion in oil, 6 mmol).After stirring for about 15 minutes at r.t., isobutyl iodide (420 □L,3.65 mmol) was added and the solution warmed to 60° C. After stirring at60° C. overnight, the reaction was quenched with saturated aqueousammonium chloride and diluted with EtOAc. The solution was washed twicewith water and brine, and then dried (Na₂SO₄), filtered andconcentrated. Purification by flash chromatography (40M Biotage column,10-30% linear gradient EtOAc/Hex, over 45 min at 35 mL/min) affordedabout 1.07 g (2.6 mmol, 87%) of the title compound as a white foam.LRMS: 412.3 (M+1)

Preparation 83A 1-Boc[2-(methyl-methanesulfonyl-amino)-phenyl]-piperazine

[0979]

[0980] The title compound was prepared in a manner similar toPreparation 82A except that methyl iodide was used and K₂CO₃ instead ofNaH as the base. LRMS (ESI+): 370.2 (M+1)

Preparation 84A1-Boc-4-[2-(ethyl-methanesulfonyl-amino)-phenyl]-piperazine

[0981]

[0982] The title compound was prepared in a manner similar toPreparation 82A except that ethyl iodide was used and K₂CO₃ instead ofNaH as the base. LRMS (ESI+): 384.2 (M+1)

Preparation 85A1-Boc-4-[2-(n-butyl-methanesulfonyl-amino)-phenyl]-piperazine

[0983]

[0984] The title compound was prepared in a manner similar toPreparation 82A except that n-butyl iodide was used. LRMS (ESI+): 412.2(M+1)

Preparation 86A1-Boc-4-{2-[(2-ethyl-butyl)-methanesulfonyl-amino]-phenyl}-piperazine

[0985]

[0986] The tide compound was prepared in a manner similar to Preparation82A except that 1-bromo-2-ethylbutane was used. LRMS (ESI+): 440.2 (M+1)

Preparation 87A1-Boc-4-[2-(cyclohexylmethyl-methanesulfonyl-amino)-phenyl]-piperazine

[0987]

[0988] The tide compound was prepared in a manner similar to Preparation82A except that bromomethyl cyclohexane was used. LRMS: 452.2(M+1)

Preparation 88A1-Boc-4-[2-(cyclobutylmethyl-methanesulfonyl-amino)-phenyl]-piperazine

[0989]

[0990] The title compound was prepared in a manner similar toPreparation 82A except that bromomethyl cyclobutane was used. LRMS(ESI+): 424.1 (M+1)

Preparation 89A1-Boc-4-[2-(cyclopropylmethyl-methanesulfonyl-amino)-phenyl]-piperazine

[0991]

[0992] The title compound was prepared in a manner similar toPreparation 82A except that bromomethyl cyclopropane was used. LRMS(ESI+): 410.1 (M+1)

Preparation 90A1-Boc-4-{2-[methanesulfonyl-(3-methyl-butyl)-amino]-phenyl}-piperazine

[0993]

[0994] The title compound was prepared in a manner similar toPreparation 82A except that 1-iodo-3-methyl butane was used. LRMS(ESI+): 426.2 (M+1)

Preparation 91A1-Boc-4-[2-(1,1-dioxo-2-isothiazolidinyl)-phenyl]-piperazine

[0995]

[0996] To a solution of N-boc-4-(2-amino-phenyl)-piperazine (555 mg, 2.0mmol) and Et₃N (837 uL, 6 mmol) in 20 mL of CH₂Cl₂ was added3-chloropropanesulfonyl chloride (255 uL, 2.1 mmol). After stirring forabout 30 minutes, the mixture was quenched with saturated aqueous sodiumbicarbonate, diluted with EtOAc, washed with 1 M HCl, water and brine,and then dried (Na₂SO₄), filtered and concentrated. Purification bysilica gel chromatography (35 g SiO₂, 10 to 30% EtOAc/hexanes, over 30min at 35 mL/min) afforded about 781 mg (1.87 mmol, 93%) ofN-boc-4-[2-(3-chloro-propane-1-sulfonylamino)-phenyl]-piperazine as awhite solid. LRMS (ESI+): 418.1 [M+1]

[0997] To a solution ofN-boc-4-[2-(3-chloro-propylamino)-phenyl]-piperazine (593 mg, 1.42 mmol)in 140 mL of DMF was added NaH (567 mg of a 60% dispersion in oil, 14mmol). After stirring for about 1 hour, the mixture was quenched withsaturated aqueous sodium bicarbonate, diluted with EtOAc, washed withwater and brine, and then dried (Na₂SO₄), filtered and concentrated toafford about 740 mg ofN-boc4-[2-(1,1-dioxo-isothiazolidin-2-yl)-phenyl]-piperazine. LRMS(ESI+): 382.1 [M+1]

Preparation 92A 1-Boc-4-(2-ethanesulfonylamino-phenyl)-piperazine

[0998]

[0999] To a solution of 1.0 g (4.4 mmol) of1-Boc-4-(2-amino-phenyl)-piperazine and 1.1 mL (6.6 mmol) oftriethylamine in 12 mL of DCM was added 0.63 mL (6.6 mmol) ofethanesulfonyl chloride, and the mixture was stirred at r.t. for about16 hours. The mixture was diluted with ethyl acetate and washed oncewith 10% aqueous sodium bisulfate and then once with saturated aqueoussodium bicarbonate. The organic portion was dried (Na₂SO₄), filtered andconcentrated in vacuo. Silica gel chromatography (3iotage, 40% ethylacetate/hexanes) of the residue afforded about 0.73 g (45%) of the titlecompound. LRMS (ESI−): 368 (M−1)

Preparation 93A 1-Boc-4-(2-n-butanesulfonylamino-phenyl)-piperazine

[1000]

[1001] The title compound was prepared in a manner similar toPreparation 92A except that n-butanesulfonyl chloride was used. LRMS(ESI+): 398 (M+1)

Preparation 94A 1-Boc-4-[2-(propane-2-sulfonylamino)-phenyl]-piperazine

[1002]

[1003] The tide compound was prepared in a manner similar to Preparation92A except that propane-2-sulfonyl chloride was used and DBU was used asthe base. LRMS (ESI+): 384.3 (M+1)

Preparation 95A 1-Boc4-(2-benzenesulfonylamino-phenyl)-piperazine

[1004]

[1005] The title compound was prepared in a manner similar toPreparation 92A except that benzenesulfonyl chloride was used. LRMS(ESI+): 418.1 (M+1)

Preparation 96A 1-Boc4-(2-Phenylmethanesulfonylamino-phenyl)piperazine

[1006]

[1007] The title compound was prepared in a manner similar toPreparation 92A except that α-toluenesulfonyl chloride was used. LRMS(ESI+): 432 (M+1)

Preparation 97A 1-Boc-(2-piperazin-1-yl-phenyl)-N,N-dimethylsulfonimide

[1008]

[1009] To a 0° C. anhydrous methylene chloride (10 mL) solution of1-Boc-4-(2-amino-phenyl)-piperazine (1.0 gm, 3.61 mmol), and TEA (0.60mL, 4.33 mmol) was added dimethylsulfamoyl chloride (0.46 mL, 4.33mmol). The bath was removed after 5 minutes, and the reaction wasstirred under a nitrogen atmosphere for about 3 days and refluxed for 1day. The mixture was diluted with methylene chloride and 1N HCl. Theseparated aqueous layer was extracted with methylene chloride (2×). Thecombined organics were dried (sodium sulfate), filtered, andconcentrated to afford crude oil. Silica gel chromatography (0 to 5%methanol in methylene chloride) gave about 0.2 g (14%) of the finalproduct. LRMS (ESI+): 385.3

Preparation 98A 1-Boc-4-[2-(acetyl-isobutyl-amino)-phenyl]-piperazine

[1010]

[1011] To a solution of N-boc-4-(2-isobutylamino-phenyl)-piperazine (333mg, 1.0 mmol, 1.0 eq), Et₃N (0.42 mL, 3.0 mmol, 3.0 eq) and DMAP (6 mg,0.05 mmol, 0.05 eq) in DCM (10 mL) was added acetic anhydride (0.14 mL,1.5 mmol, 1.5 eq). The mixture was stirred at r.t. overnight. Thereaction was diluted with DCM (50 mL) and washed with saturated aqueousNaHCO₃ (25 mL) and brine (25 mL). The organic layer was separated andaqueous layer was extracted with DCM (2×). The combined organic extractswere dried (Na₂SO₄), filtered and concentrated to afford the titlecompound (375 mg, 1.0 mmol, 100%). LRMS (ESI+): 376.18 (M+1)

Preparation 99A1-Boc-4-[2-(isobutyl-methoxycarbonyl-amino)-phenyl]-piperazine

[1012]

[1013] The title compound was prepared in a manner similar toPreparation 98A except that methyl chloroformate was used instead ofacetic anhydride. LRMS (ESI+): 392.2 (M+1)

Preparation 100A1-Boc-4-[2-(isobutyl-isopropoxycarbonyl-amino)-phenyl]-piperazine

[1014]

[1015] The title compound was prepared in a manner similar toPreparation 98A except that isopropyl chloroformate was used instead ofacetic anhydride. LRMS (ESI+): 420.26 (M+1).

Preparation 101A1-Boc-4-[2-(isobutyl-isobutoxycarbonyl-amino)-phenyl]-piperazine

[1016]

[1017] The title compound was prepared in a manner similar toPreparation 98A except that isobutyl chloroformate was used instead ofacetic anhydride. LRMS (ESI+): 434.27 (M+1).

Preparation 102A1-Boc-4-{2-[(2,2-dimethyl-propoxycarbonyl)-isobutyl-amino]-phenyl}-piperazine

[1018]

[1019] The title compound was prepared in a manner similar toPreparation 98A except that neopentyl chloroformate was used instead ofacetic anhydride. LRMS (ESI+): 448.32 (M+1).

Preparation 103A4-{2-[(1-methyl-1H-imidazol-ylmethyl)-amino]-phenyl}piperazine

[1020]

[1021] To a solution of 1-boc-4-(2-aminophenyl) piperazine (554 mg, 2.0mmol, 1.0 eq.) in methanol was added(1-methyl-1H-imidazole-2-carbaldehyde (220 mg, 2.0 mmol, 1.0 eq.). Themixture was reflux for about 1 hour and then cool to about 0° C. Sodiumborohydride on alumina (10 wt % on basic alumina, 1.13 g, 3.0 mmol, 1.5eq.) was added. The solution was warmed to r.t. and then stirredovernight. The reaction mixture was filtered through celite and thenconcentrated. Purification by flash chromatography (35 g SiO₂, 40mL/min, linear gradient, 0-8% MeOH/CH₂Cl₂ for 25 minutes and then 8%MeOH for 7 minutes) afforded Boc protected title compound (176 mg, 0.47mmol, 24%). LRMS (ESI+): 372.3 [M+1].

Preparation 104A 2-(N-Boc-piperazin-1-yl)-benzaldehyde

[1022]

[1023] To a solution of 1-(2-cyanophenyl)-piperazine (375 mg, 2.0 mmol)in 15 mL of dioxane was added DIBAL-H (6 mL of a 1 M solution inheptane, 6 mmol). After stirring at r.t. for about 48 hours, thesolution was transferred via cannula into 20 mL of 0.5 M Rochelle saltAfter stirring for about 2 hours, NaHCO₃ (636 mg, 6 mmol) and Boc₂O (567mg, 2.6 mmol) were added. After stirring overnight, EtOAc and brine wereadded. After separation, the aqueous solution was extracted with EtOAc(3×). The combined organic layers were washed with water and brine, andthen dried (Na₂SO₄), filtered and concentrated. Purification by flashchromatography (35 g SiO₂, linear gradient 10-20% EtOAc/Hex over 30 minat 35 mL/min) afforded about 436 mg (1.50 mmol, 75%) of the titlecompound as a yellow oil. LRMS (ESI+): 291.1 (M+1)

Preparation 105A 1-Boc-4-(2-pyrrolidin-1-ylmethyl-phenyl)-piperazine

[1024]

[1025] To a solution of 2-(N-Boc-piperazin-1-yl)-benzaldehyde (400 mg,1.4 mmol) in pyrrolidine (0.33 mL, 4 mmol) was added titaniumisopropoxide (1.2 mL, 4 mmol), and the mixture was stirred at r.t. undera nitrogen atmosphere. After about 30 minutes, the mixture was dilutedwith ethanol (4 mL). Sodium borohydride (106 mg, 2.8 mmol) was added andthe mixture was stirred for about 16 hours. Water (2 mL) was added, andthe resulting suspension was filtered. The filter cake was washed withmethanol (5 mL), and the filtrate was concentrated to dryness.Purification by flash chromatography (1:1 hexanes/ethyl acetate) gavethe title compound (470 mg, 96%) as a colorless oil.

[1026]¹H NMR (CDCl₃) δ7.40-7.44 (m, 1H), 7.19-7.26 (m, 1H), 7.01-7.08(m, 2H), 3.68 (s, 2H), 3.55 (t, J=4.5 Hz, 4H), 2.92-2.95 (m, 4H), 2.53(m, 4H), 1.75 (m, 4H), 1.49 (s, 9H). TLC (SiO₂): R_(f)=0.28 (50%EtOAc/hexanes).

Preparation 106A 1-Boc-4-(2-piperidin-1-ylmethyl-phenyl)-piperazine

[1027]

[1028] The title compound was prepared in the same manner as describedin Preparation 105A except that piperidine was used.

[1029]¹H NMR (CDCl₃) δ7.38 (d, J=7.6 Hz, 1H), 7.20-7.26 (m, 1H),7.03-7.08 (m, 2H), 3.54-3.57 (m, 4H), 3.50 (s, 2H), 2.92-2.95 (m, 4H),2.40 (m, 4H), 1.23-1.59 (m, 15H). TLC (SiO₂): R_(f)=0.52 (50%EtOAc/hexanes)

Preparation 107A 1-Boc-4-(2-diethylaminomethyl-phenyl)-piperazine

[1030]

[1031] The title compound was prepared in the same manner as describedin Preparation 105A except that diethyl amine was used.

[1032]¹H NMR (CDCl₃) δ7.54 (d, J=7.3 Hz, 1H), 7.19-7.26 (m, 1H),7.03-7.11 (m, 2H), 3.63 (s, 2H), 3.56 (t, J=4.4 Hz, 4H), 2.88 (t, J=4.6Hz, 4H), 2.54 (q, J=7.2 Hz, 4H), 1.49 (s, 9H), 1.03 (t, J=7.2 Hz, 6H).TLC (SiO₂): R_(f)=0.36 (50% EtOAc/hexanes)

Preparation 108A 1-Boc-4-(2-di-n-butylaminomethyl-phenyl)-piperazine

[1033]

[1034] The title compound was prepared in the same manner as describedin Preparation 105A except that dibutyl amine was used.

[1035]¹H NMR (CDCl₃) δ7.54-7.57 (m, 1H), 7.01-7.26 (m, 3H), 3.58 (s,2H), 3.49-3.53 (m, 4H), 2.85-2.90 (m, 4H), 2.38 (t, J=7.3 Hz, 4H),1.40-1.50 (m, 13H), 0.84 (t, J=7.3 Hz, 6H). TLC (SiO₂): R_(f)=0.70 (80%EtOAc/hexanes).

Preparation 109A

[1036] 1-Boc-4-(2-morpholin-4-ylmethyl-phenyl)-piperazine

[1037] To a solution of 1-Boc-4-(2-formyl-phenyl)-piperazine (500 mg,1.7 mmol) in methanol (10 mL) was added morpholine (348 mg, 4.0 mmol)and sodium cyanoborohydride (315 mg, 5 mmol) and the mixture was stirredfor about 24 hours. The mixture was diluted with ethyl acetate (100 mL),washed with saturated NaHCO₃ solution (10 ml), water (10 mL) and brine(10 mL). The organic phase was dried over anhydrous sodium sulfate andconcentrated. Silica gel chromatography (1:1 hexanes/ethyl acetate)afforded the title compound as a colorless oil (182 mg, 30%).

[1038]¹H NMR (CDCl₃) δ7.38-7.40 (m, 1H), 7.21-7.27 (m, 1H), 7.04-7.09(m, 2H), 3.67 (t, J=4.4 Hz, 4H), 3.53-3.55 (s, 6H), 2.92-2.95 (m, 4H),2.46-2.49 (m, 4H), 1.49 (s, 9H). TLC (SiO₂): R_(f)=0.44 (50%EtOAc/hexanes).

Preparation 110A 1-Boc-4-[2-(isopropylamino-methyl)-phenyl]-piperazine

[1039]

[1040] The title compound was prepared in the same manner as describedin Preparation 105A except that isopropylamine was used.

[1041]¹H NMR (CDCl₃) δ7.30-7.00 (m, 4H), 3.80 (s, 2H), 3.60-3.45 (m,4H), 2.95-2.85 (m, 4H), 2.85-2.80 (m, 1H), 1.50 (m, 9H), 1.10-1.00 (m,6H). TLC (SiO₂): R_(f)=0.10 (25% ethyl acetate/hexanes).

Preparation 111A1-Boc-4-{2-[(Acetyl-isopropyl-amino)-methyl]-phenyl}-piperazine

[1042]

[1043] 1-Boc-4-[2-(isopropylamino-methyl)-phenyl]-piperazine (0.325 g,0.975 mmol) was dissolved in tetrahydrofuran (5 mL) and cooled to about0° C. TEA (0.54 mL, 3.9 mmol) was added followed by dropwise addition ofacetyl chloride (0.2 mL, 2.93 mmol). The solution was allowed to warm tor.t. The solvents were removed under reduced pressure, and the resultingoil was purified using silica chromatography (ethyl acetate) to affordthe title compound (0.650 g, 82%) as an oil.

[1044]¹H NMR (CDCl₃) δ7.25-6.95 (m, 4H), 4.65 (s, 1H), 4.45 (s, 1H),3.70-3.50 (m, 4H), 2.90-2.80 (m, 4H), 1.50 (s, 9H), 1.30-1.20 (m, 1H),1.10-1.00 (m, 6H).

Preparation 112A1-Boc-4-[2-(isopropyl-methanesulfonyl-amino-methyl)-phenyl]-piperazine

[1045]

[1046] The title compound was prepared in the same manner as describedin Preparation 111A except that methanesulfonyl chloride was usedinstead of acetyl chloride.

[1047]¹H NMR (CDCl₃) δ7.70-7.60 (m, 1H), 7.25-6.95 (m, 3H), 4.45 (s,2H), 3.65 (s, 2H), 3.65 (s, 3H), 2.95-2.80 (m, 8H), 1.50 (s, 9H),1.30-1.15 (m, 1H), 1.10-1.00 (m, 6H).

Preparation 113A1-Boc-4-{2-[Hydroxy-(1-methyl-1H-imidazol-2-yl)-methyl]-phenyl}-piperazine

[1048]

[1049] To a solution of 1-methyl imidazole (350 uL, 4.4 mmol) in 15 mLof THF at −78° C. was added n-BuLi (1.5 mL of a 1.6 M solution inhexane, 2.4 mmol). After stirring for about 30 minutes, the solution waswarmed to about 0° C. and then stirred for about 15 minutes. The mixturewas then cooled to about −78° C. A solution ofN-Boc-4-(2-formylphenyl)-piperazine (580 mg, 1.0 mmol) in 5 mL of THFwas added via cannula The solution was allowed to warm slowly to r.t.overnight. After addition of saturated aqueous NH₄Cl and brine, thesolution was extracted with EtOAc (2×). The combined organic solutionswere dried (Na₂SO₄), filtered and concentrated. Purification by silicagel chromatography (35 g SiO₂, 0 to 10% 0.2 M NH₃ in MeOH/CH₂Cl₂ over 30min at 35 mL/min) afforded about 592 mg (1.59 mmol, 79%) of the alcoholas a colorless oil. LRMS (ESI+): 373.2 (M+1)

Preparation 114A1-Boc-4-[2-(1-Methyl-1H-imidazole-2-carbonyl)-phenyl]-piperazine and1-Boc-4-{2-[acetoxy-(1H-imidazol-2-yl)-methyl]-phenyl}-piperazine

[1050]

[1051] To a solution of1-Boc-4-{2-[hydroxy-(1H-imidazol-2-yl)-methyl]-phenyl}-piperazine (200mg, 0.734 mmol) and sodium bicarbonate (185 mg, 2.2 mmol) in 8 mL ofCH₂Cl₂ was added Dess-Martin periodane (467 mg, 1.1 mmol). Afterstirring for about 1 hour, 2 mL of saturated aqueous sodium bicarbonateand 2 mL 0.5 M Na₂S₂O₃ were added. After stirring for about 1 hour, thesolution was diluted with CH₂Cl₂ and washed with water and brine, andthen dried (Na₂SO₄), filtered and concentrated. Purification by silicagel flash chromatography (35 g SiO₂, 0-5% 0.2 M NH₃ in MeOH/CH₂Cl₂linear gradient over 30 min at 35 mL/min) afforded about 53 mg (0.14mmol, 19%) of the ketone {LRMS (ESI+): 371.2 [M+1]} and about 114 mg(0.28 mmol, 37%) of the acetate {LRMS (ESI+): 415.2 [M+1]}.

Preparation 115A1-Boc-4-[2-(1-methyl-1H-imidazol-2-ylmethyl)-phenyl]-piperazine

[1052]

[1053] To a solution of1-Boc-4-{2-[hydroxy-(1H-imidazol-2-yl)-methyl]-phenyl}-piperazine (93mg, 0.25 mmol) in 5 mL of THF was added NaH (30 mg, 0.75 mmol). Afterstirring for about 45 minutes, CS₂ (75 uL, 1.25 mmol) was added. Afterstirring for about 30 minutes, 5 mL of THF was added followed by MeI (78uL, 1.25 mmol). After stirring for about 1 hour, saturated aqueous NH₄Cland brine were added. The solution was extracted EtOAc (2×). Thecombined organic solutions were dried (Na₂SO₄), filtered andconcentrated. Purification by silica gel chromatography (35 g SiO₂, 0 to5% MeOH/CH₂Cl₂ over 30 minutes at 35 mL/min) afforded about 97 mg (0.21mmol, 84%) of the xanthate as a yellow oil. LRMS (ESI+): 463.2 (M+1)

[1054] To a solution of the xanthate (90 mg, 0.195 mmol) and Bu₃SnH (260uL, 0.967 mmol) in 2 mL of toluene at 80° C. was added AIBN (50 uL of a0.4 M solution in toluene, 0.02 mmol). Another 50 uL of the AIBNsolution was added every 2 to 3 hours for 8 hours. After stirringovernight another 50 uL of the AIBN solution was added. After stirringfor about 8 more hours, the solution was concentrated and filteredthrough celite with CH₂Cl₂. Purification by silica gel chromatography(35 g SiO₂, 0 to 5% 0.2 M NH₃ in MeOH/CH₂Cl₂ over 30 minutes at 35mL/min) afforded about 46 mg (0.13 mmol, 66%) of the deoxygenatedproduct as a colorless oil. LRMS (ESI+): 357.2 (M+1)

Preparation 116A 1-Boc-4-(2-thiazol-2-ylmethyl-phenyl)-piperazine

[1055]

[1056] Thiazole was lithiated with n-butyllithium and reacted withN-Boc-4-(2-formylphenyl)-piperazine in a manner similar to preparation113A. The resulting alcohol was deoxygenated in a manner similar topreparation 115A to afford the final compound. LRMS (ESI+): 360.1 (M+1)

Preparation 117A1-Boc-4-[2-(2-methyl-2H-[1,2,4]triazol-3-ylmethyl)-phenyl]-piperazine

[1057]

[1058] 1-methyltetrazole was lithiated with n-butyllithium and reactedwith N-Boc-4-(2-formylphenyl)-piperazine in a manner similar topreparation 113A. The resulting alcohol was deoxygenated in a mannersimilar to preparation 115A to afford the final compound. LRMS (ESI+):358.3 (M+1)

Preparation 118A 1-Boc-4-(2-isobutoxy-phenyl)-piperazine

[1059]

[1060] To a solution of 1-Boc-4-(2-hydroxy-phenyl)-piperazine (560 mg,2.0 mmol) in 10 mL of DMF was added K₂CO₃ (835 mg, 6 mmol). Afterstirring for about 5 minutes, isobutyl iodide (350 uL, 3 mmol) wasadded. After stirring overnight at 60° C., the solution was warmed toabout 80° C. After stirring for about 4 hours, the solution was cooledto r.t. diluted with EtOAc, washed with water and brine, and then dried(Na₂SO₄), filtered and concentrated. Purification by silica gelchromatography (35 g SiO₂, 10 to 30% EtOAc/hexanes, over 45 minutes at35 mL/min) afforded about 418 mg (1.25 mmol, 62%) ofN-Boc-4-(2-isobutoxy-phenyl)-piperazine as a colorless oil. LRMS (ESI+):335.1 [M+1].

Preparation 119A1-Boc-4-[2-(1-methyl-1H-imidazol-2-ylmethoxy)-phenyl]piperazine

[1061]

[1062] To a solution of 1-Boc-4-(2-hydroxy-phenyl)-piperazine (556 mg,2.0 mmol, 1.0 eq.), (1-methyl-1H-imidazol-2-yl)-methanol (448 mg, 4.0mmol, 2.0 eq.), triphenylphosphine (1.04 g mg, 4.0 mmol, 2.0 eq.) andTHF at 0° C. under nitrogen was added DEAD (0.629 mL, 4.0 mmol, 2.0 eq.)slowly so that temperature of the reaction does not rise above 10° C.After addition was complete, the ice bath was removed and the mixturewas stirred at r.t. overnight. Methanol was added, and the mixture wasstirred for about 15 minutes and then concentrated. Purification byflash chromatography (35 g SiO₂, 40 mL/min, linear gradient 0-8% 2.0 MNH₃ in MeOH/CH₂Cl₂ for 25 minutes and 8% 2.0 M NH₃ in MeOH for 7minutes) afforded the title compound (279 mg, 0.75 mmol, 37%). LRMS(ESI+): 373.3 (M+1).

Preparation 120A 1-Boc-4-(2-benzyloxy-phenyl)-piperazine

[1063]

[1064] The title compound was prepared in a manner similar toPreparation 118A except that benzyl bromide was used. LRMS (ESI+): 369.1(M+1).

Preparation 121A 1-Boc-4-(2-carboxy-phenyl)-piperazine

[1065]

[1066] To a solution of 1-(2-cyanophenyl)-piperazine (37.45 g, 200 mmol)in 500 mL of absolute ethanol was added 1000 mL of 25% aqueous KOH. Thesolution was heated to reflux for about 72 hours and then cooled toabout 0° C. The solution was acidified with 890 mL of 5 M HCl, and thensolid NaHCO₃ was added to bring the pH of the solution to about 8.NaHCO₃ (12.7 g, 120 mmol) and Boc₂O (11.4 g, 52.2 mmol) were added andthe mixture was stirred overnight, which was then acidified with 5 M HClto about pH 1. After addition of EtOAc and brine, the aqueous solutionwas separated and extracted with EtOAc (2×). The combined organicsolutions were washed with water (2×) and brine, and then dried(Na₂SO₄), filtered, and concentrated. The material was purified byrecrystallization from EtOAc/hexanes to afford about 49.8 g (162 mmol,81%) of the title compound. LRMS (ESI−): 305.2 (M−1)

Preparation 122A (2-piperazin-1-yl-phenyl)-piperadin-1-yl-methanone

[1067]

[1068] The 1-Boc-4-(2-carboxy-phenyl)-piperazine (1 g, 3.26 mmol),piperidine (278 mg, 3.26 mmol), EDCI (625 mg, 3.26 mmol) and DMAP (50mg, catalytic) were dissolved in DCM (20 mL) and stirred at r.t. forabout 12 hours. The mixture was washed with water, dried, filtered, andconcentrated. The resultant foam was taken up in DCM (10 mL) and TPA (5mL) was added, and the mixture stirred at r.t. for about 2 hours. Thereaction was concentrated and subjected to SCX ion exchangechromatography followed by silica gel chromatography to afford the finalproduct (868 mg, 71%) as a white foam. LRMS (ESI+): 274.1 (M+1).

Preparation 123A 1-Boc-4-[2-(2H-tetrazol-5-yl)-phenyl]-piperazine

[1069]

[1070] A solution of 4-(2-cyano-phenyl)-piperazine (1.7 g, 9.0 mmol, 1.0eq) in azidotributyltin (5.0 g, 15 mmol, 1.5 eq.) was stirred at 80° C.for about 5 days. Purification by SCX (10 g) ion exchange chromatographyafforded crude 4-(2-tetrazole-5-yl-phenyl)-piperazine. LRMS (ESI+):231.0 (M+1).

[1071] To a solution of 4-[2-(2H-tetrazole-5-yl-phenyl)-piperazine 1.8g, 7.7 mmol, 1.0 eq.), NaHCO₃ (978 mg, 9.2 mmol, 1.2 eq), DMAP (94 mg,0.77 mmol, 0.1 eq.) in H₂O:Dioxane (1:1) was added (Boc)2O (1.6 g, 7.7mmol, 1.0 eq.). The resulting mixture was stirred at r.t. overnight, andthen neutralized with 1.0 M HCl and extracted with EtOAc (3×). Thecombined organic extracts were dried (Na₂SO₄), filtered andconcentrated. Purification by flash chromatography (35 g SiO₂, lineargradient, 40 mL/min , 0%-10% 2.0 M NH₃ in MeOH/CH₂Cl₂ for 20 minutes and10% 2.0 M NH₃ in MeOH /CH₂Cl₂ for 13 minutes) afforded the titlecompound (798 mg, 2.41 mmol, 32%). LRMS (ESI+): 331.1 (M+1).

Preparation 124A 4-[2-(2-isobutyl-2H-tetrazol-5-yl)-phenyl]piperazineand 4-[2-(1-isobutyl-1H-tetrazol-5-phenyl]piperazine

[1072]

[1073] To a solution of N-boc-4-[2-(2H-tetrazole-5-yl-phenyl)-piperazine(330 mg, 1.0 mmol, 1.0 eq.) in DMF (10 mL) was added K₂CO₃ (331 mg, 2.4mmol, 2.4 eq.) and isobutyl iodide (0.14 mL, 1.2 mmol, 1.2 eq.). Themixture was stirred at r.t. overnight. The mixture is diluted with ethylacetate (50 mL) and washed with H₂O (20 mL) and brine (20 mL). Theaqueous layers were extracted with EtOAc (2×). The combined organicextracts were dried (Na₂SO₄), filtered and concentrated to afford about370 mg (96%, 0.96 mmol) of a mixture (60:40 by NMR) of Boc-protectedtitle compounds favoring the 2H substituted tetrazole. LRMS (ESI+):387.2 (M+1).

[1074] To a solution of the mixture of Boc-protected compounds fromabove (360 mg, 0.93 mmol, 1.0 eq.) in CH₂Cl₂ (10 mL) was added TPA (5mL) and DMS (0.25 mL). The resulting mixture was stirred at r.t. forabout 2 hours. The reaction mixture was concentrated and purified usingSCX (10 g) ion-exchange chromatography to afford a mixture of the titlecompounds (240 mg, 0.84 mmol, 90%). LRMS (ESI+): 287.1 (M+1).

[1075] C Domain Preparations:

[1076] The protected amino acid derivatives corresponding to the B and Cdomains are, in many cases, commercially available. Other protectedamino acid derivatives can be prepared by following known literaturemethods (See Williams, R. M. Synthesis of Optically Active α-AminoAcids, Pergamon Press: Oxford, 1989). The following provides thepreparation of C domains.

Preparation 1C1-Methoxycarbonylmethyl-1,3-dihydro-isoindole-2-carboxylic acidtert-butyl ester

[1077]

[1078] Step A: (2-Bromo-benzyl)-carbamic acid tert-butyl ester

[1079] To a mixture of 125.0 g (561.8 mmol) of 2-bromobenzylaminehydrochloride and 170.7 g (1236.0 mmol) of potassium carbonate in 300 mLof 50% THF/water was added 134.9 g (618.0 mmol) of di-tert-butyldicarbonate in four portions over 20 minutes. The mixture was stirred atr.t. for about 16 hours and then diluted with 300 mL of ethyl acetateand 300 mL of water. The organic portion was separated and the aqueousportion was extracted three times with 200 mL each of ethyl acetate. Thecombined ethyl acetate portions were washed once with 250 mL of 10%aqueous sodium bisulfate. The organic portion was dried (MgSO₄),filtered and concentrated to dryness to afford about 161 g of Step Acompound.

[1080] Step B: 3-[2-(tert-Butoxycarbonylamino-methyl)-phenyl]-acrylicacid methyl ester

[1081] To compound of Step A (161.0 g, 561.8 mmol) in DMF (800 mL) wasadded methyl acrylate (58.0 g, 674.2 mmol), TEA (170.5 g, 1685.4 mmol)and dichlorobis(triphenylphosphine) palladium(II) (7.9 g, 11.2 mmol).The mixture was heated at 80° C. for about 32 hours. The mixture wascooled, diluted with 1000 mL of EtOAc and washed with 10% aqueous sodiumbisulfate. The aqueous portion was extracted three times with EtOAc andthe combined organics were dried (Na₂SO₄) and concentrated to dryness.The residue was dissolved in a small amount of DCM and filtered through7 inches of silica gel in a 2 L sintered glass funnel eluting with 25%EtOAc/hexanes. The eluent was concentrated to dryness and recrystallizedfrom EtOAc/hexanes to afford about 116.9 g (71%) of Step B compound.

[1082] Step C: To a 0° C. solution of (116.9 g, 401.2 mmol) materialfrom Step B in DCM (800 mL) was added 200 mL of TFA dropwise over 15minutes. After removing the cooling bath, the mixture was stirred forabout 2.5 hours and then concentrated to dryness. The residue wasdissolved in 500 mL of DCM and saturated aqueous sodium bicarbonate isslowly added until the mixture was slightly basic. The organic portionwas separated and the aqueous portion is extracted two times with DCM.The combined organic portions were dried (Na₂SO₄) and concentrated todryness. The residue was dissolved in 800 mL of DCM and DIPEA (57.0 g,441.4 mmol) was added. To the mixture was added di-tert-butyldicarbonate (96.3 g, 441.4 mmol) in five portions over 45 minutes andthen stirred at r.t. for 16 hours. The mixture was washed with 10%aqueous sodium bisulfate, and the organic portion was separated and theaqueous portion is extracted two times with DCM. The combined organicextracts were dried ((Na₂SO₄) and concentrated to dryness. The resultingresidue was dissolved in a small amount of DCM and filtered through 7inch silica gel in a 2L sintered glass funnel eluting with 25%EtOAc/hexanes. The eluent was concentrated to dryness and theenantiomers were separated by chiral chromatography. The first elutingisomer was labeled as isomer #1 and the second eluting is labeled asisomer #2, which afforded about 52.6 g (45%) of the final compound(isomer 2). EIS-MS 292 [M+1].

Preparation 2C 1-Carboxymethyl-1,3-dihydro-isoindole-2-carboxylic acidtert-butyl ester

[1083]

[1084] To 1-methoxycarbonylmethyl-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester (52.6 g, 180.5 mmol) in MeOH (500 mL) was added 1N NaOH (199 mL, 199.0 mmol). The mixture is stirred at r.t. for about 48hours and then concentrated to dryness. The resulting residue wasdissolved in water (300 mL) and extracted with diethyl ether (2×). Theaqueous portion was acidified to pH 2 with 10% aqueous sodium bisulfateand extracted with EtOAc. The combined organic extracts were dried(MgSO₄) and concentrated to dryness to afford about 49.8 g of the finalcompound (99%). EIS-MS 276 [M−1].

Preparation 3C (2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid

[1085]

[1086] Step A: (2,3-dihydro-1H-isoindol-1-yl)-acetic acid methyl ester:

[1087] To the compound prepared in Preparation C1 (11.75 g., 40.41 mmol)in DCM (50 mL) was added TFA (50 mL) dropwise. After about 2 hours, themixture was concentrated to dryness and the resulting residue waspartitioned with saturated aqueous sodium bicarbonate (200 mL) and EtOAc(300 mL). The organic portion was separated and the aqueous layer wasextracted with DCM (4×500 mL). The combined DCM extracts were combined,dried (Na₂SO₄), and concentrated to dryness to afford about 3.97 g(51%).

[1088] Step B: (2-isopropyl-2.3-dihydro-1H-isoindol-1-yl)-acetic acidmethyl ester:

[1089] To the compound obtained from Step A (0.50 g, 2.61 mmol) indichloroethane (46 mL) was added acetone (1.76 mL, 24.01 mmol) andsodium triacetoxyborohydride (2.48 g., 11.74 mmol). After 6 hours, themixture was diluted with 1.0N NaOH (100 mL), and the organic portion wasseparated. The aqueous layer was extracted with DCM (3×100 mL). Thecombined DCM extracts were dried (MgSO₄) and concentrated to dryness toafford about 0.60 g (99%). EIS-MS 235 [M+1].

[1090] Step C: To the compound of Step B (0.53 g., 2.30 mmol) in MeOH(5.1 mL) was added 1.0N NaOH (2.53 mL, 2.53 mmol). After two days, thesolution was concentrated to dryness. The resulting residue was dilutedwith 1.0N HCl and water was loaded onto a strong cation exchange resin.The resin was washed with water, TBF/water (1:1) and then water. Theproduct was then eluted from the resin with pyridine/water (1:9). Theeluent was concentrated to dryness to afford about 0.43 g (85%) of thefinal compound. EIS-MS 220 [M+1].

Preparation 4C (2-Methyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid

[1091]

[1092] Step A: (2-Methyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acidmethyl ester:

[1093] The compound from preparation C1 was deprotected with TFA in amanner similar to preparation 3C of Step A. To the deprotected compound(0.50 g, 2.61 mmol), in dichloroethane (46 mL), was added 37% aqueousformaldehyde solution (1.80 mL, 24.01 mmol) and sodiumtriacetoxyborohydride (2.48 g., 11.74 mmol). After 3 days, the mixturewas diluted with 1.0N NaOH (100 mL). The organic portion was separatedand the aqueous layer was extracted with DCM (3×100 mL). The combinedDCM extracts were dried (Na₂SO₄) and concentrated to dryness. Theresulting residue was purified by flash chromatography (SiO₂, elutingwith 100% EtOAc) affording about 0.43 g (79%) of the alkylatedisoindole. EIS-MS 206 [M+1].

[1094] Step B: To the compound of Step A (0.34 g., 1.66 mmol) in MeOH(3.7 mL) was added 1.0N NaOH (1.82 mL, 1.82 mmol). After 2 days, thesolution was concentrated to dryness. The resulting residue was dilutedwith 1.0N HCl and water was then loaded onto a strong cation exchangeresin. The resin was washed with water, THF/water(1:1) and water, andthe product was eluted from the resin with pyridine/water(1:9). Theeluent was concentrated to dryness to afford about 0.31 g (98%) of thefinal compound. EIS-MS 192 [M+1].

Preparation 5C

[1095]

[1096] The above compound was prepared from Boc-L-Tic-OH as described inPreparation 6C below, except that the Weinreb amide was made by asimilar procedure to that described in Synthesis, 676, 1983.

Preparation 6C

[1097]

[1098] Boc-D-Tic-OH (14.9 g, 53.7 mmol), methoxymethylaminehydrochloride (5.24 g, 53.7 mmol), EDC (11.3 g, 59.1 mmol), HOBT (7.98g, 59.1 mmol), DIEA (9.83 ml, 59.1 mmol) and THF (500 ml) were combined,and the resulting mixture was stirred for about 18 hours at r.t. undernitrogen. The reaction mixture was concentrated and the residue wastaken up in ethyl acetate. The resulting mixture was washed with 1M HCl,saturated NaHCO₃ and brine, which was then dried via filtration throughphase separator paper. Removal of solvent gives a residue, which waschromatographed on silica gel using (1:1 ethylacetate /hexane) to giveabout 12.3 g of Boc-D-Tic-NMeOMe (Weinreb amide).

[1099] Lithium aluminum hydride (1.0M in THF, 5.1 ml, 5.00 mmol) wasslowly added to the Weinreb amide prepared above (1.28 g, 4.00mmol) inTHF (35 ml) at 0° C. The reaction mixture was stirred at 0° C. for about15 minutes. Aqueous KHSO₄ (970 mg in 20 ml H₂O) was slowly addedfollowed by diethylether. The organic layer was separated and theaqueous layer was extracted with diethylether. The organic phases werecombined and washed with aqueous 1M HCl, saturated aqueous NaHCO₃ andbrine, which was then dried over Na₂SO₄. Removal of solvent affordedabout 780 mg of the final compound. MS: MH+262.

Preparation 7C (2-Butyl-2.3-dihydro-1H-isoindol-1-yl)-acetic acid methylester

[1100]

[1101] The compound from preparation C1 was deprotected with TFA in amanner similar to preparation 3C of Step A. To the deprotected compound(0.50 g, 2.61 mmol) and butryaldehyde (2.16 mL, 24.01 mmol) indichloroethane (46 mL) was added sodium triacetoxyborohydride (2.48 g.,11.74 mmol). After reacting about 3 hours, the mixture was diluted with1.0 N NaOH (100 mL) and partitioned. The aqueous layer was extractedwith DCM (3×75 mL). The DCM layers were combined, dried over sodiumsulfate, filtered and concentrated under reduced pressure to give abrown residue. The residue was purified via silica gel chromatography(eluent: ethyl acetate/hexanes (1:3). The purified fractions werecombined and concentrated to give the title compound as a brown oil(0.51 g, 77%). MS ES 249.2 (M+H)

Preparation 8C (2-Butyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid

[1102]

[1103] To a solution containing the compound 7C (0.47 g, 1.89 mmol) inmethanol (4.2 mL) was added 1.0 N NaOH (2.08 mL, 2.08 mmol). Afterreacting about 2 hours, the solution was concentrated under reducedpressure. The residue was diluted with 1.0 N HCl, and water was loadedonto a strong cation exchange resin. The resin was washed with water andTHF/water (1:1), and the product was eluted from the resin withpyridine/water (1:9). The pyridine washes were concentrated underreduced pressure, and azeotroped with acetone to give the tide compoundas brown solids (0.28 g., (64%)) MS ES 234.19 (M+H)

Preparation 9C

[1104]

[1105] Step A: To a solution of N-Boc-4-Fluoro-D-Phe (2.37 g, 8.366mmol) in methanol, 3 mL of concentrated sulfuric acid was added. Themixture was heated to reflux overnight and then concentrated in vacuo.MS M+1 198.1

[1106] Step B: To an ice cold mixture of 1.65 g (8.367 mmol) of compoundfrom Step A, 1.353 mL of pyridine and ethyl chloroformate (0.848 mL,8.869 mmol) is added slowly with stirring for about 30 minutes givingwhite solid. The mixture was partitioned between water and ethylacetate. The aqueous layer was extracted with EtOAc (2×). The combinedorganic solution was dried over MgSO₄, filtered, and concentrated invacuo to give about 2.17 g of yellow oil (96%). MS M+1 270.1.

[1107] Step C: A mixture containing 2.17 g (8.06 mmol) of the compoundfrom Step B, paraformaldehyde (0.254 g, 8.46 mmol), and 10 mL of 3:1glacial acetic acid/conc. sulfuric acid was stirred at r.t. for about 48hours. The mixture was partitioned between water and ethyl acetate. Theaqueous layer was extracted with EtOAc (3×). The combined EtOAc solutionwas dried over magnesium sulfate, filtered, and concentrated in vacuo.The desired product was purified by column chromatography eluting with25% EtOAc in Hexane to give about 1.31 g (58%) of colorless oil. MS: M+1282.1

[1108] Step D: A solution of 1.31 g (4.656 mmol) of material from Step Cin 20 mL of 5N HCl was heated to reflux for about 24 hours. The solutionwas concentrated in vacuo. The resulting white solid was washed withether to afford about 0.87 g (81%). MS M+1 196.1.

[1109] Step E: To a solution of 0.87 g (3.755 mmol) of material fromStep D in 20 ml of 1:1 dioxane/water, di-t-butyl-dicarbonate (0.901 g,4.131 mmol) and 2.355 mL (16.90 mmol) of TEA were added. The mixture wasallowed to stir at r.t. overnight. The mixture was diluted with EtOAc,and the separated aqueous layer was extracted with EtOAc (3×). Thecombined organic solution was dried over magnesium sulfate, filtered andconcentrated in vacuo to give about 0.64 g (58%) of the final compound.MS M-1 294.1.

Preparation 10C

[1110]

[1111] Step A: By following a procedure of Preparation 28C, Step A and1.0 g (5.58 mmol) of □-methyl-DL-phenylanaline, about 1.4 g of ester wasprepared. MS M+1 194.1

[1112] Step B: By following a procedure of Preparation 28C, Step B and1.08 g (5.59 mmol) of material from Step A, about 1.48 g (100%) ofproduct was prepared. MS M+1 266.1

[1113] Step C: By following a procedure of Preparation 28C, Step C and1.48 g (5.59 mmol) of material from Step B, about 1.55 g (100%) ofproduct was prepared. MS M+1 278.1

[1114] Step D: By following a procedure of Preparation 28C, Step D and1.55 g (5.59 mmol) of material from Step C, about 1.33 g of product wasprepared. MS M+1192.1

[1115] Step E: By following a procedure of Preparation 28C, Step E and1.33 g (5.84 mmol) of material from Step D, about 1.70 g (100%) of thefinal compound was prepared. MS M+1 292.2

[1116] Preparation 11C

[1117] Step A: By following a procedure of Preparation 28C, Step A and2.0 g (11.16 mmol) of □-methyl-D-phenylanaline, about 2.15 g of esterwas prepared. MS M+1 194.1

[1118] Step B: By following a procedure of Preparation 28C, Step B and2.15 g (11.16 mmol) of material from Step A, about 1.46 g (49%) ofproduct was prepared. MS M+1 266.1

[1119] Step C: By following a procedure of Preparation 28C, Step C and1.46 g (5.503 mmol) of material from Step B, about 0.74 g (48%) ofproduct was prepared. MS M+1 278.1

[1120] Step D: By following a procedure of Preparation 28C, Step D and0.74 g (2.67 mmol) of material from Step C, about 0.54 g (89%) ofproduct was prepared. MS M+1192.1

[1121] Step E: By following a procedure of Preparation 28C, Step E and0.54 g (2.37 mmol) of material from Step D, about 0.54 g (78%) of thefinal compound was prepared. MS M+1 292.2

Preparation 12C

[1122]

[1123] Step A: By following a procedure of Preparation 28C, Step A and0.65 g (1.95 mmol) of N-Boc-4-trifluoromethyl-D-phenylanaline, about0.48 g of ester was prepared. MS M+1 248.0

[1124] Step B: By following a procedure of Preparation 28C, Step B and0.48 g (1.95 mmol) of material from Step A, about 0.60 g (96%) ofproduct was prepared. MS M+1 320.1

[1125] Step C: By following a procedure of Preparation 28C, Step C and0.6 g (1.879 mmol) of material from Step B, about 0.37 g (59%) ofproduct was prepared. MS M+1 332.1

[1126] Step D: By following a procedure of Preparation 28C, Step D and0.37 g (1.117 mmol) of material from Step C, about 0.11 g (35%) ofproduct was prepared. MS M+1 246.1

[1127] Step E: By following a procedure of Preparation 28C, Step E and1.11 g (0.391 mmol) of material from Step D, about 0.234 g (>100%) ofthe final compound is prepared. MS M−1 344.1

Preparation 13C Lithium;(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetate

[1128]

[1129] Step 1: (1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetic acid methylester

[1130] To a solution 100.4 g (52 mol) of Boc-tetrahydoisoquinoline-1-acetic (100.4 g 520.0 mmol) in 200 mL methanol was added400 mL of 2.3 M HCl in methanol. The mixture was stirred overnight andconcentrated in vacuo. The residue was dissolved in ethyl acetate andwashed with saturated sodium bicarbonate, brine, and then dried (Na₂SO₄)and concentrated in vacuo to afford about 109.5 g (100%) of the titlecompound. EIS-MS: 206 (M+1).

[1131] Step 2:1-methoxycarbonylmethyl-3,4-dihydro-1H-isoguinoline-2-carboxylic acidtert-butyl ester

[1132] To a 0° C. solution of material from Step 1 (50.5 g, 240.0 mmol)in 250 mL dry THF was added di-tert-butyl dicarbonate (59.3 g, 270.0mmol) in 50 mL dropwise. After stirring for about 45 minutes, themixture was concentrated in vacuo. The residue was dissolved in ethylacetate, washed with saturated sodium bicarbonate and brine, and thendried (Na₂SO₄) and concentrated in vacuo. Chromatography of the residueafforded both enantiomers of the title compound.

[1133] EIS-MS: 306 (M+1).

[1134] Step 3: To a solution of material from Step 2 (10.2 g, 33.4 mmol)in 220 mL of dioxane was added a solution of lithium hydroxidemonohydrate (1.67 g, 39.8 mmol) in 110 mL water in portions to maintaina temperature below 30° C. The mixture was stirred for about 16 hoursand concentrated in vacuo to afford about 11.2 g of the final compound.

[1135] EIS-MS: 292 (M+1).

Preparation 14C Lithium;(2-methyl-1,2,3,4-tetrahydro-isoquinolin-1-yl)-acetate

[1136]

[1137] Step 1: (1,2,3,4-Tetrahydro-isoquinolin-1-yl)-acetic acid methylester

[1138] The material from Preparation of 13C Step 2 (9.98 g, 32.7 mmol)was mixed with 500 mL cold 4M HCl/dioxane and stirred at r.t. for aboutan hour. The mixture was concentrated in vacuo. The residue wasdissolved in ethyl acetate and then washed with saturated sodiumbicarbonate and brine. The organic portion was dried (Na₂SO₄), filteredand concentrated in vacuo to afford about 6.9 g (100%) of the titlecompound. EIS-MS: 206 (M+1).

[1139] Step 2: (2-methyl-1,2,3,4-tetrahydro-isoquinolin-1-yl)-aceticacid methyl ester

[1140] To a solution of material from Step 1 (6.71 g, 32.0 mmol) in 175mL of dichloroethane was added 37% aqueous formaldehyde (22.6 mL, 300mmol). After about 10 minute, sodium triacetoxyborohydride (31.2 g,147.0 mmol) was added in 2 to 3 g portions with some cooling to maintainambient temperature. The mixture was stirred for about 16 hours and DCMand water was added. The mixture was adjusted to pH 9-10 with 5N sodiumhydroxide. The organic layer was separated, washed with brine, and thendried (Na₂SO₄) and concentrated in vacuo. Chromatography (silica gel, 5%(2N ammonia in methanol)/DCM) of the residue afforded about 6.9 g (96%)of the title compound. EIS-MS: 220 (M+1).

[1141] Step 3: To a solution of material from Step 2 (4.45 g, 18.9 mmol)in 120 mL dioxane was added lithium hydroxide monohydrate (1.02 g, 22.7mmol) in 65 mL water in portions keeping the temperature below 30° C.After about 16 hours, the mixture was concentrated in vacuo to affordabout 8.12 g of the final compound. EIS-MS: 206 (M+1).

Preparation 15C1,1-Dimethyl-6-methoxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acidethyl ester

[1142]

[1143] To a solution of the triflate salt of1,1-dimethyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid ethylester (1.5 g, 3.76 mmol, 1.0 eq.) in MeOH (20 mL) and CH₂Cl₂ (2 mL) at0° C. was added a solution of (trimethylsilyl)diazomethane (2.0 M inhexane, 3.7 mL, 2.0 eq.). The resulting mixture was warmed to r.t. andstirred overnight, and then the solution was concentrated. Purificationby flash chromatography (125 g SiO₂ linear gradient, 40 mL/min, 1:1EtOAc/hexane for 33 minutes) afforded about 900 mg of the final compound(96%). LRMS (electrospray): 250.2 (M+1).

[1144] “A Domain” and “B Domain” Combination

Preparation 1AB and 2AB1-(D-p-Cl-Phe)-4-(2-methanesulfonyl-phenyl)piperazine and1-(D-p-Cl-Phe)-4-(2-methanesulfinyl-phenyl)-piperazine

[1145]

[1146] Commercially available 1-(2-methylthiophenyl) piperazine wascoupled to Boc-p-Cl-D-Phe-OH in a manner substantially similar to thatdescribed in coupling procedure 1. To a solution of the coupled product(100 mg, 0.204 mmol) in 5 mL of CH₂Cl₂ cooled to −78° C. was addedm-chloroperbenzoic acid (49 mg, 0.204 mmol). After stirring for about 30minutes, the reaction was quenched with 1 M Na₂S₂O₃ and extracted withCH₂Cl₂. The combined organic solutions were washed with saturated sodiumbicarbonate, dried (Na₂SO₄), filtered and concentrated. Purification byflash chromatography (10 g SiO₂, linear gradient 0-10% methanol/CH₂Cl₂,30 mL/minute, over 30 minutes) afforded about 46 mg (0.090 mmol, 43%) ofthe sulfoxide and 60 mg (0.115 mmol, 56%) of the sulfone. Each of theseis separately deprotected in manner substantially similar to thatdescribed in coupling procedure 1.

Preparation 3AB

[1147]

[1148] 1-(2-Nitrophenyl)piperazine (3.13 g, 15.1 mmol),Boc-D-4-chlorophenylalanine (4.52 g, 15.1 mmol), EDC (3.19 g, 16.6mmol), HOBT (2.21 g, 16.7 mmol) and DIBA (2.63 ml, 15.1 mmol) were addedto THF. The resulting mixture was stirred overnight at r.t. under anitrogen. The reaction mixture was then concentrated in vacuo. Theresidue was taken up in ethyl acetate and washed with 1M HCl, dilutedwith NaHCO₃ and brine and then dried with Na₂SO₄. Removal of the solventgave a residue, which was chromatographed on normal phase (ethylacetate/hexane 1:1) to give about 6.8 g of the Boc-protected compound.

[1149] The Boc-protected compound (6.88 g, 14.1 mmol) was dissolved in4M HCl/dioxane (230 ml), and the resulting mixture was stirred at r.t.for about an hour. The mixture was concentrated in vacuo to give about5.1 g of the final compound.

Preparation 4AB

[1150]

[1151] The mixture of piperazine from Preparation 54A (6.99 g, 28.76mmol), N-Boc-D-Cl-Phe (8.624 g, 28.76 mmol), HATU (10.94 g, 28.76 mmol)and DIEA (25.05 mL, 143.8 mmol) in 160 mL of DCM was stirred at r.t.overnight. The mixture was partitioned between water and CH₂Cl₂. Theaqueous layer was extracted with CH₂Cl₂ (2×). The combined organicsolution was dried over MgSO₄, filtered and concentrated in vacuo. Theresidue was Purified through silica gel column using 10% MeOH in EtOActo give Boc protected product. The Boc protected compound was treatedwith 1:1 TFA/CH₂Cl₂. The mixture was stirred at r.t. for about 2 hoursand then concentrated in vacuo to afford the final compound (13.9 g,74%). MS M+1 425.2.

Preparation 5AB 1-(D-p-Cl-Phe)-4-1-[(2-aminosulfonyl)phenyl]piperazine

[1152]

[1153] 1-[(2-aminosulfonyl)phenyl]piperazine from preparation 19A wascoupled to Boc-p-Cl-D-Phe-OH followed by deprotection and HCl saltformation in a manner similar to coupling procedure 1, Steps 1 and 4.

[1154] “B Domain” and “C Domain” Combination

Preparation 1BC N-Boc-D-Tic-D-p-Cl-phe-OH

[1155]

[1156] Step 1: The HCl salt of H-D-p-Cl-Phe-OMe (35.8 g, 129 mmol) wasdissolved in water (200 mL). Ethyl acetate (200 mL) was added followedby addition of a saturated sodium bicarbonate solution. The mixture wasstirred for about 5 minutes, and then the organic layer was separated,washed with water (200 mL) and dried over magnesium sulfate.Concentration of the mixture under reduced pressure produces a whitesolid (32.2 g). The solid was then dissolved in methylene chloride (200mL), D-Boc-Tic (35.8 g, 129 mmol) and 4-dimethylaminopyridine (75 mg).The mixture was cooled to 0° C. and EDC (24.7 g, 129 mmol) was added intwo portions. After stirring for about 20 minutes, the ice bath wasremoved and the solution was allowed to warm to r.t. The solution wasstirred for about 4 hours and then diluted with water (400 mL). Theorganic layer was washed with water (3×), dried over magnesium sulfateand concentrated under reduced pressure to give a clear oil (70 g).Column chromatography (35% ethyl acetate/heptane) afforded about 55.6 gof the intermediate Boc-D-Tic-D-p-Cl-Phe-OMe (85%).

[1157]¹H NMR(DMSO) (Two rotomers observed) δ8.26(d, 1H), 8.19(d, 0.5 H),7.24(d, 2H), 7.00-7.19(m, 8H), 4.68(m, 0.5H), 4.20-4.60(m, 4.5H),3.58(s, 3H), 3.51(s, 1.5H), 2.77-3.10(m, 6H), 1.42(s, 3H), 1.21(s, 9H).MS(ES) 473.0(M⁺), 471.1(M⁻).

[1158] Step 2: The compound of Step 1 (54.3 g, 114 mmol) was dissolvedin methanol (170 mL). The solution was cooled to 0° C. with an ice bathand 1N NaOH (290 mL) is added dropwise. After vigorous stirring forabout 20 minutes, the mixture was warmed to about 25° C. The solutionwas concentrated under reduced pressure to give yellow oil. The oil wasdissolved in water (200 mL) and the pH is adjusted to about 1. Ethylacetate (200 mL) was added, and the organic layer was separated anddried over magnesium sulfate. Concentration of the organics producedabout 46.3 g of the final compound.

[1159] 1H NMR(DMSO) (Two rotomers observed) δ7.98(d, 1H), 7.82(d, 0.5H), 6.90-7.41(m, 16H), 4.20-4.70(m, 8.5H), 2.60-3.20(m, 8.5H),1.32-1.41(m, 19H). MS(ES) 459.1 m/z(M⁺), 457.1(M⁻).

Preparation 2BC

[1160]

[1161] The above compound was prepared using N-Boc-L-Tic-OH as describedin Preparation 1BC.

[1162]¹H NMR(DMSO) (Two rotomers observed) δ7.98(d, 1H), 7.72(d, 0.5 H),6.90-7.41(m, 16H), 4.0-4.70(m, 8.5H), 2.60-3.20(m, 8.5H), 1.32-1.41(m,19H). MS (ES) 459.1 m/z(M⁺), 457.1(M⁻).

Preparation 3BC Lithium;2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-3-(4-chloro-phenyl)-propionate

[1163]

[1164] Step A:3-{[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylamino]-methyl}-3,4-dihydro-1H-isoguinoline-2-carboxylicacid tert-butyl ester

[1165] To a 0° C. solution of 4-Cl-D-Phe-OMe (6.27 g, 25.1 mmol) andsodium acetate (8.23 g, 100.0 mmol), in 850 ml dry MeOH, was added thealdehyde from Prearation 6C (9.8 g, 37.6 mmol) in 50 ml MeOH. Themixture was stirred for about 15 minutes and then sodiumcyanoborohydride (2.37 g, 37.6 mmol) was added. The cooling bath wasremoved and the reaction stirred for 16 hours at r.t. The mixture wasconcentrated to dryness and the resulting residue taken up in water and1 ml of 1M HCl. The mixture was extracted with EtOAc, and the organicswere washed with saturated sodium bicarbonate and brine, and then dried(Na₂SO₄) and concentrated to dryness. The resulting residue was purifiedby flash chromatography (SiO₂, eluting with 2:1 hexane/EtOAc) affordingabout 8.62 g (75%). EIS-MS 459 [M+1].

[1166] Step B: To a 12° C. solution of material from Step A (1.11 g,2.42 mmol) in dioxane (15 ml) was added a solution of lithium hydroxide(0.10 g, 2.42 mmol) in water (7.5 mL). The mixture was stirred for about16 hours and then concentrated to dryness affording about 1.08 g (100%)of the final compound. EIS-MS 445 [M+1].

Preparation 4BC Lithium;2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-3-(4-chloro-phenyl)-propionate

[1167]

[1168] The above compound was Prepared in a manner similar to thepreparation 3BC above except that aldehyde from Preparation 5C was used.

Preparation 5BC Preparation of Lithium2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-methyl-amino]-3-(4-chloro-phenyl)-propionate

[1169]

[1170] Step A: To a solution of3-{[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylamino]-methyl}-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester from preparation 3BC Step A (0.60 gm, 1.31 mmol)in anhydrous methanol, was added sodium acetate (0.54 gm, 6.54 mmol).The solution was brought to pH 5-6 with 3-4 drops of glacial aceticacid. Aqueous formaldehyde (37% by wt., 0.49 mL) was added. The solutionwas put under a nitrogen atmosphere and cooled to 0° C. After about 15minutes, sodium cyanoborohydride (0.25 gm, 3.92 mmol) was added andrinsed into the reaction with anhydrous methanol (5 mL). The mixture wasstirred at r.t. overnight, and then concentrated in vacuo andreconstituted in aqueous sodium bicarbonate and ethyl acetate. Afterseparation of phases, the aqueous phase was extracted with ethyl acetate(2×), and all organics were combined, dried (magnesium sulfate),filtered, and concentrated to an opaque white oil (0.64 gm).Chromatography (0 to 20% ethyl acetate in hexane) gave about 0.6 g ofmethylated product as a clear oil (97%). MS (m/z, ES+): 473.2.

[1171] Step B: A solution of LiOH.H₂O (0.05 gm, 1.27 mmol) in distilledwater (4 mL) was added to a solution of the material from Step A in1,4-dioxane (8 mL), and the reaction was cooled slightly in an ice waterbath. The mixture was stirred under a nitrogen atmosphere at r.t.overnight. An additional 1.5 eq. of LiOH.H₂O (0.08 gm) were added as anaqueous solution (4 mL), and the mixture was stirred at r.t. over theweekend. The mixture was concentrated, and then combined with THF andconcentrated (3×) to help dry the material. The resulting foam was driedat r.t. overnight in a vacuum oven to give about 0.67 g of finalcompound as a white foam (114%). MS (m/z, ES+): 459.2.

Preparation 6BC Lithium2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-(2-methoxy-ethyl)-amino]-3-(4-chloro-phenyl)-propionate

[1172]

[1173] Step A: To a solution of methoxyacetaldehyde (0.15 gm, 2.03mmol),3-{[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethylamino]-methyl}-3,4dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester from preparation 3BC Step C (0.31 gm, 0.68 mmol)in acetonitrile was added sodium triacetoxyborohydride (0.72 gm, 3.38mmol). After stirring overnight under a nitrogen atmosphere at r.t.,additional acetaldehyde (0.25 gm) dissolved in acetonitrile and sodiumtriacetoxyborohydride (0.21 gm) was added, and the mixture was stirredfor about 8.5 hours. The mixture was quenched at r.t. with 5N NaOH (5mL). The aqueous phase was separated from the organic and extracted withethyl acetate (4×). The combined organics were washed with a brinesolution, and then dried, filtered and concentrated. Chromatography(gradient of ethyl acetate in hexane, 0 to 12%) gives about 0.23 g of3-{[[2-(4-Chloro-phenyl)-1-methoxycarbonyl-ethyl]-(2-methoxy-ethyl)-amino]-methyl}-3,4dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester as a yellow oil (70%). MS (m/z, ES+): 517.2.

[1174] Step B: To a solution of the material from Step A in 1,4-dioxanewas added a solution of lithium hydroxide monohydrate (0.05 gm, 1.11mmol) in distilled water (2 mL). The mixture was stirred overnight atr.t. and then concentrated to a white residue. Addition of THF andconcentration (3×) gives the lithium carboxylate as a foam. The foam wasdried overnight under vacuum to afford about 0.25 g of crude solids(109%). MS (m/z, ES+): 503.3.

Preparation 7BC1-{[1-Carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-methyl}-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester

[1175]

[1176] Step A: To a suspension of 4-Cl-D-Phe-OMe hydrochloride(40.4 g,161.5 mmol) in DCM (250 mL) was added saturated aqueous sodiumbicarbonate (250 mL), and the mixture was stirred at r.t. for about 1hour. The organic portion was separated and the aqueous portion wasextracted with DCM (2×). The combined organic portions were dried(Na₂SO₄) and concentrated to dryness. To the free amine, in DCM (400 mL)at 0° C., was added 1-Carboxymethyl-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester from preparation 2C (isomer 2, 44.8 g, 161.5mmol), EDC (31.0 g, 161.5 mmol) and 4-DMAP (2.0 g, 16.1 mmol). Themixture was stirred at 0° C. for about 30 minutes whereupon the coolingbath was removed and the mixture was stirred for another 5 hours at r.t.The mixture was then washed with saturated aqueous sodium bicarbonate(200 mL) and 10% aqueous sodium bisulfate (200 mL), and then dried(Na₂SO₄) and concentrated to dryness to afford about 76.4 g (100%) ofthe ester. EIS-MS 471 [M−1].

[1177] Step B: To the ester from Step A (76.4 g, 161.5 mmol) in MeOH(760 mL) was added 1 N NaOH (242.0 mL, 242.0 mmol), and the mixture washeated at 50° C. for 4 hours and then stirred for another 16 hours atr.t. After concentrating to dryness, the resulting residue was taken upin 500 mL of water and washed with diethyl ether (2×). The aqueousportion was acidified to pH 2 with 10% aqueous sodium bisulfate andextracted with EtOAc (4×200 mL). The combined organic extracts weredried (MgSO₄) and concentrated to dryness. The resulting solid wassuspended in hexanes, filtered, and dried to afford about 67.7 g (91%)of the final compound. EIS-MS 457 [M−1].

Preparation 8BC3-(4-Chloro-phenyl)-2-[(1,1-dimethyl-1,2,3,4-tetrahydro-isoquinoline-3-carbonyl)-amino]-propionicacid methyl ester

[1178]

[1179] To a solution of 1,1-dimethyl Tic (240 mg, 1.17 mmol),4-Cl-D-Phe-OMe (322 mg, 1.28 mmol), HOBT (197 mg, 1.46 mmol), and DIPEA(0.81 mL, 44.68 mmol) in DCM/DMF (1:1) was added EDC (280 mg, 1.46mmol). The resulting mixture was stirred at r.t. overnight. The mixturewas then diluted with EtOAc (100 mL), washed with saturated aqueousNaHCO₃ and brine, and then dried Na₂SO₄) and concentrated to dryness.Purification and separation of diastereomers by flash chromatography (35g SiO₂, linear gradient, 40 mL/min 10-50% EtOAc/hexane for 25 minutesand 50% EtOAc/hexane for 7 minutes) afforded the final compound. LRMS(ESI+): 401.1 (M+H).

Preparation 9BC3-(4-Chloro-phenyl)-2-[(1,1-dimethyl-1,2,3,4-tetrahydro-isoquinoline-3-carbonyl)-amino]-propionicacid

[1180]

[1181] To the compound from preparation 8BC (5.95 g, 14.88 mmol) in a1:1 mixture of THF/H₂O (50 mL) was added lithium hydroxide hydrate (0.75g, 17.87 mmol). The mixture was stirred at r.t. for about 18 hours. Themixture was then concentrated to dryness. The resulting residue wasdissolved in water (50 mL), made acidic with 1N HCl (25 mL) and washedwith Et₂O (100 mL). The aqueous layer was evaporated to dryness toafford about 6.18 g of the final compound (98%). LRMS(EIS+): 387 [M+1].

Preparation 10BC1-{[1-Carboxy-2-(4-methoxy-phenyl)-ethylcarbamoyl]-methyl}-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester

[1182]

[1183] Step 1: To a solution of p-methoxy-D-Phe-OMe (1.72 g, 8.23 mmol)dissolved in TBF (45mL) and1-carboxymethyl-1,3-dihydro-isoindole-2-carboxylic acid tert-butyl ester(2.51 g, 9.05 mmol) was added HOBT (1.22 g, 9.05 mmol), EDC (1.73 g,9.05 mmol) and DIPEA (1.6 mL, 9.05 mmol). The reaction was stirredovernight at r.t. and then concentrated. The mixture was washed with 1MHCl, dilute NaHCO₃ and brine, and then dried with sodium sulfate. Themixture was chromatographed on silica gel eluting with 3% 2M NH₃ inMeOH/CH₂Cl₂ giving about 2.58 g as white solids. Mass MH⁺ 469

[1184] Step 2: The white solid from Step 1 (2.58 g, 5.5 mmol) wasdissolved in dioxane (37 mL) and lithium hydroxide hydrate (0.35 g, 8.3mmol) dissolved in H₂O (19 mL) was added. The mixture was stirred forabout 2.5 hours at r.t. and then concentrated. Ethyl acetate was addedand the mixture was treated with 1M HCl, which was then washed withbrine and concentrated to afford about 2.56 g of the final free acid.LRMS(ESI+): 455 (M+1)

Preparation 11BC1-[1-Carboxy-2-(4-chloro-phenyl)-ethylcarbamoyl]-1,3-dihydro-isoindole-2-carboxylicacid tert-butyl ester

[1185]

[1186] Step 1: About 2.0 g (7.60 mmol) of (R,S)-Boc-1,3-dihydro-2Hisoindole carboxylic acid was dissolved in 100 ml THF and about 2.28 g(9.12 mmol) of 4-Cl-D-phe-methylester HCl, 1.25 g (9.12 mmol) of HOBT,1.75 g (9.12 mmol) of EDC, and 1.6 ml (9.12 mmol) of DIEA were added.The mixture was stirred overnight at r.t., concentrated to dryness,washed with 1M HCl, dilute NaHCO₃ and brine, and then dried over sodiumsulfate. The material was chromatographed on silica gel by eluting withethyl acetate/hexane 1:2 to give about 1.05 g of isomer 1 and about 0.82g of isomer 2, and about 1.61 g mixture of isomers 1 and 2. Mass MH⁺459.

[1187] Step 2: About 0.82 g (1.79 mmol) of the isomer 2 obtained in Step1 was dissolved in 11 ml of dioxane and 0.11 g (2.68 mmole) ofLiOH-hydrate in 5.5 ml of H₂O was added. The mixture was stirred forabout 4 hours at r.t. and then concentrated to dryness. Ethyl acetatewas added, and the solution was washed with 1M HCl and brine, and thenconcentrated to dryness affording about 0.75 g of the free acid. Mass:445 (MH⁺).

EXAMPLE Coupling Procedure 11-(D-TIC-4-Cl-D-Phe)-4-(2-methylphenyl)-piperazine, HCl

[1188]

[1189] Step 1: To a solution of N-Boc-4-Cl-D-Phe (200 mg, 0.67 mmol, 1.0eq.), 1-(2-methylphenyl)piperazine (140 mg, 0.79 mmol, 1.2 eq.), HOBT(113 mg, 0.84 mmol, 1.25 eq.), DIPEA (290 microliters, 1.66 mmol, 2.5eq.), CH₂Cl₂ (4 mL), and DMP (1 mL) is added EDC (160 mg, 0.84 mmol,1.25 eq.). The solution is stirred at r.t. overnight. The solution isdiluted with ethyl acetate (30 mL) and washed with saturated aqueoussodium bicarbonate, 0.05 M phosphate buffer (pH 7, 2×) and brine, andthen dried over Na₂SO₄, filtered, and concentrated. Purification byflash chromatography (10 g SiO₂, linear gradient 0-10% methanol/CH₂Cl₂,30 mL/minute, over 15 minutes) affords about 293 mg (96%) of1-(N-Boc-4-Cl-D-Phe)-4-(2-methylphenyl)piperazine. LRMS (ESI+): 458.2(M+H).

[1190] Step 2: To a solution of1-(N-Boc-4-Cl-D-Phe)-4-(2-methylphenyl)piperazine (293 mg, 0.64 mmol),CH₂Cl₂ (2 mL), and DMS (0.5 mL) is added TFA (2 mL). After stirring forabout 1 hour, the solution is azeotroped from heptane (3×). The residueis dissolved in CH₂Cl₂ and washed with saturated sodium bicarbonate. Theaqueous solution is extracted with CH₂Cl₂ (2×). The combined organicsolutions were dried over Na₂SO₄, filtered, and concentrated to affordabout 200 mg (87%) of 1-(4-Cl-D-Phe)-4-(2-methylphenyl)piperazine.

[1191] Step 3: To a solution of1-(4-Cl-D-Phe)-4-(2-methylphenyl)piperazine (60 mg, 0.17 mmol, 1.0 eq.),N-Boc-D-TIC (56 mg, 0.20 mmol, 1.2 eq.), HOBT (28 mg, 0.21 mmol, 1.25eq.), DIPEA (73 microliters, 0.42 mmol, 2.5 eq.), CH₂Cl₂ (2 mL) andDMF(0.5 mL) is added EDC (40 mg, 0.21 mmol, 1.25 eq.). The solution isstirred at r.t. overnight. The solution is diluted with ethyl acetateand washed with saturated aqueous sodium bicarbonate, 1 M NaHSO₄ andbrine, and then dried over Na₂SO₄, filtered and concentrated.Purification by flash chromatography (10 g SiO₂, linear gradient 0 to100% EtOAc/CH₂Cl₂, 30 mL/min, over 30 minutes) gives about 81 mg (0.13mmol, 77%) of 1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-methylphenyl)piperazine.LRMS (ESI+): 617.2 (M+H).

[1192] Step4: To a solution of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-methylphenyl)piperazine (81 mg, 0.13mmol), CH₂Cl₂ (2 mL), and DMS (0.5 mL) is added TFA (2 mL). Afterstirring for about 1 hour, the solution is azeotroped from heptane (3×).The residue is dissolved in CH₂Cl₂ and washed with saturated aqueoussodium bicarbonate. The aqueous solution is extracted with CH₂Cl₂ (2×).The combined organic solutions are dried over Na₂SO₄, filtered andconcentrated. The residue is dissolved in 5% methanol/Et₂O andprecipitated with 1 M HCl in Et₂O. The precipitate is washed with Et₂O(2×) to afford about 64 mg (0.12 mmol, 92%) of the title compound. HRMS(ESI+) calculated for C₃₀H₃₄ClN₄O₂: 517.2370. Found: 517.2383 (M+H).

Coupling Procedure 21-(D-TIC-4-Cl-D-Phe)-4-(2-methoxy-5-nitrophenyl)piperazine, HCl

[1193]

[1194] Step 1: To a solution of N-Boc-D-TIC-4-Cl-D-Phe-OH (348 mg, 0.76mmol, 1.2 eq.), 1-(2-methoxy-5-nitrophenyl]piperazine (150 mg, 0.63mmol, 1.0 eq.), HOAT (108 mg, 0.79 mmol, 1.25 eq), 2,6-lutidine (0.37mL, 3.18 mmol, 5.0 eq.), CH₂Cl₂ (8 mL) and DMF (2 mL) is added HATU (300mg, 0.79 mmol, 1.25 eq.). After stirring at r.t. overnight, the solutionis diluted with ethyl acetate and washed with 1 M HCl (2×), saturatedsodium bicarbonate and brine, and then dried over Na₂SO₄, filtered, andconcentrated. Purification by flash chromatography (10 g SiO₂, lineargradient 0-5% methanol/CH₂Cl₂, 30 mL/minute, over 20 minutes) givesabout 392 mg (0.58 mmol, 91%) of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-methoxy-5-nitrophenyl) piperazine. LRMS(ESI+): 678.5 (M+H).

[1195] Step 2: To a solution of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-methoxy-5-nitrophenyl)piperazine (53 mg,0.078 mmol) in CH₂Cl₂ (2 mL) and DMS (0.2 mL) is added TFA (1 mL). Afterstirring for about 2 hours, the solution is azeotroped from heptane(2×). The residue is dissolved in CH₂Cl₂ and washed with saturatedsodium bicarbonate. The aqueous solution is extracted with CH₂Cl₂ (3×).The combined organic solutions are dried over Na₂SO₄, filtered, andconcentrated. Purification by flash chromatography (10 g SiO₂, lineargradient 0-10% methanol/CH₂Cl₂, 30 mL/minute, over 30 minutes) affordsD-TIC-4-Cl-D-Phe-4-(2-methoxy-5-nitrophenyl)piperazine. The solid isdissolved in CH₂Cl₂ and precipitated with 1 M HCl in Et₂O. The solutionis concentrated to afford about 40 mg (0.065 mmol, 84%) of the titlecompound. HRMS (ESI+) calculated for C₃₀H₃₄ClN₄O₂: 578.2170. Found:578.2157 (M+H).

Coupling Procedure 31-(D-TIC-4-Cl-D-Phe)-4-(2-methanesulfonyl-phenyl)piperazine, HCl

[1196]

[1197] Step 1. To a solution of1-(D-p-Cl-Phe)-4-(2-methanesulfinyl-phenyl)-piperazine (Preparation1AB)(168 mg, 0.39 mmol, 1.0 eq.), N-Boc-D-TIC (132 mg, 0.47 mmol, 1.2eq.), HOBT (69 mg, 0.49 mmol, 1.25 eq.), DIPEA (0.17 mL, 1.0 mmol, 2.5eq.), CH₂Cl₂ (5 mL) and DMF (2 mL) is added EDC (95 mg, 0.49 mmol, 1.25eq.). The solution is stirred at r.t. overnight. The solution is dilutedwith EtOAc and washed with saturated aqueous NaHCO₃ and brine, and thendried over Na₂SO₄, filtered and concentrated. Purification by flashchromatography (35 g SiO₂, 40 mL/min, linear gradient, 40-60%EtOAc/hexane over 15 min and 60% EtOAC/Hexane for 18 minutes) affords(256 mg, 0.39 mmol, 96%)1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-methanesulfonyl-phenyl)piperazine. LRMS(ESI+): 681.2 (M+H).

[1198] Step 2. To a solution of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-methanesulfonyl-phenyl)piperazine (240mg, 0.35 mmol), CH₂Cl₂ (2 mL) and DMS (0.25 mL) is added TFA (2 mL).After stirring for about 2 hours, the solution is azeotroped fromheptane (3×). The residue is dissolved in CH₂Cl₂ and washed withsaturated sodium bicarbonate. The aqueous layer is extracted with CH₂Cl₂(2×). The combined organic extracts are dried (Na₂SO₄), filtered, andconcentrated. The residue are dissolved in 5% MeOH/Et₂O and precipitatedwith 1 M HCl in Et₂O. The precipitates are washed with Et₂O (2×) toafford (191 mg, 0.31 mmol, 88%) the chloride salt of title compound.HRMS (ESI+) calcd. for C₃₀H₃₄ClSN₄O₄: 581.1989. Found: 581.1995.

Coupling Procedure 41-(D-TIC-4-Cl-D-Phe)-1-(5-isopropyl-2-pyrrolidin-1-ylmethyl-phenyl)-piperazine,3HCl

[1199]

[1200] Step 1:1-Boc-4-(5-isopropyl-2-pyrrolidin-1-ylmethyl-phenyl)-piperazine (162 mg,0.42 mmol, 1.0 eq.) is deprotected with TFA and freebased using SCX ionexchange chromatography. To a solution of the deprotected piperazine,the BC domain from Preparation 3BC (245 mg, 0.54 mmol, 1.3 eq.), HOBT(68 mg, 0.50 mmol, 1.2 eq.), Et₃N (140 microliters, 1.0 mmol, 2.4 eq.),CH₂Cl₂ (4 mL) and DMF (4 mL) is added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (96 mg, 0.5mmol, 1.2 eq). The solution is stirred at r.t. overnight. The solutionis diluted with ethyl acetate (30 mL) and washed with saturated sodiumbicarbonate, water and brine, and then dried over Na₂SO₄, filtered andconcentrated. Purification by flash chromatography (35 g SiO₂, lineargradient 0-10% 2 M NH₃ in methanol/CH₂Cl₂, 35 mL/minute, over 30minutes) affords about 250 mg (0.35 mmol, 84%) of2-Boc-3-({1-(4-chloro-benzyl)-2-[4-(5-isopropyl-2-pyrrolidin-1-ylmethyl-phenyl)-piperazin-1-yl]-2-oxo-ethylamino}-methyl)-3,4-dihydro-1H-isoquinoline.LRMS (ESI+): 714.2 (M+H).

[1201] Step 2: To a solution of the compound from Step 1 (240 mg, 0.035mmol) in CH₂Cl₂ (2 mL) and DMS (0.2 mL) is added TFA (1 mL). Afterstirring for about 2 hours, the solution is azeotroped from heptane(2×). The residue is dissolved in CH₂Cl₂ and washed with saturatedsodium bicarbonate. The aqueous solution is extracted with CH₂Cl₂ (3×).The combined organic solutions are dried over Na₂SO₄, filtered, andconcentrated. Purification by flash chromatography (35 g SiO₂, lineargradient 0-10% 2M NH₃ in methanol/CH₂Cl₂, 35 mL/minute, over 30 minutes)affords the title compound. The solid is dissolved in CH₂Cl₂ andprecipitated with 1 M HCl in Et₂O. The solution is concentrated toafford about 235 mg (0.325 mmol, 93%) of the title compound. HRMS (ESI+)calculated for C₃₇H₄₉ClN₅O: 614.3626. Found: 614.3627 (M+H).

Coupling Procedure 5N-{1-(4-Chloro-benzyl)-2-oxo-2-[4-(2-pyrrolidin-1-ylmethyl-phenyl)-piperazin-1-yl]-ethyl}-2-(2-methyl-2,3-dihydro-1H-isoindol-1-yl)-acetamide,2HCl

[1202]

[1203] To a room temperature, stirred solution of2-amino-3-(4-chlorophenyl)-1-[4-((2-pyrrolidin-1-yl)methylphenyl)-piperazin-1-yl]-propan-1-one(0.49g., 1.15 mmol), (2-methyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid(0.17 g., 0.1.15 mmol) and HATU (0.43 g., 1.15 mmol) in DCM is addedN,N-diisopropylethylamine (0.40 mL, 2.31 mmol). After about an hour, thesolution is concentrated under reduced pressure, and the residue ispurified by silica gel chromatography (eluent: 5 to 10% 2.0 M NH₃ inMeOH)/DCM). The purified fractions are combined and concentrated to giveBoc protected compound as a yellow film (0.15 g, 22%) LRMS (ESI+): 600.2(M+H)

[1204] To a flask containing(R)-N-{1-(4-chlorobenzyl)-2-oxo-2-[4-((2-pyrrolidin-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-2-(2-methyl-2,3-dihydro-1H-isoindol-1-yl)-acetamide is added 1.0 N HCl(7 mL). After about 1 hour, the solution is solidified at −78° C. andthe solid is lyophilized to give the title compound as purple solids(0.10 g.) LRMS (ESI+): 600.2 (M+H)

Examples 1-83

[1205] The compounds of Examples 1-83 are prepared from an appropriate Adomain piperazine by following a substantially similar couplingprocedure as described in Procedures 1-5.

Exam- Coupling ple Z Procedure MS (ESI) 1

1 582.2 (M + H) 2

1 548.2 (M + H) 3

1 616.2 (M + H) 4

2 567.2 (M + H) 5

2 562.2 (M + H) 6

1 517.2 (M + H) 7

1 548.2 (M + H) 8

1 571.2 (M + H) 9

1 531.3 (M + H) 10

1 549.2 (M + H) 11

2 531.5 (M + H) 12

1 531.3 (M + H) 13

1 528.2 (M + H) 14

2 528.0 (M + H) 15

2 519.2 (M + H) 16

2 521.2 (M + H) 17

2 537.2 (M + H) 18

2 521.2 (M + H) 19

2 517.2 (M + H) 20

2 531.3 (M + H) 21

2 545.3 (M + H) 22

2 559.0 (M + H) 23

1 531.3 (M + H) 24

1 533.2 (M + H) 25

1 547.2 (M + H) 26

2 561.3 (M + H) 27

2 575.3 (M + H) 28

3 565.2 (M + H) 29

3 581.2 (M + H) 30

2 577.1 (M + 1) 31

2 563.2 (M + 1) 32

2 563.3 (M + 1) 33

2 563.3 (M + 1) 34

2 601.3 (M + 1) 35

2 595.2 (M + 1) 36

2 602.2 (M + 1) 37

2 533.2 (M + 1) 38

2 584.3 (M + 1) 39

2 585.2 (M + 1) 40

2 583.3 (M + 1) 41

2 629.3 (M + 1) 42

2 629.3 (M + 1) 43

2 586.1 (M + 1) 44

2 600.0 (M + 1) 45

2 602.0 (M + 1) 46

2 597.3 (M + 1) 47

2 625.3 (M + 1) 48

2 611.3 (M + 1) 49

2 643.3 (M + 1) 50

2 642.0 (M + 1) 51

2 640.0 (M + 1) 52

2 560.2 (M + 1) 53

2 594.0 (M + 1) 54

2 588.0 (M + 1) 55

2 616.0 (M + 1) 56

2 610.0 (M + 1) 57

2 624.0 (M + 1) 58

2 638.0 (M + 1) 59

2 672.0 (M + 1) 60

2 624.0 (M + 1) 61

2 638.0 (M + 1) 62

2 700.0 (M + 1) 63

2 575.0 (M + 1) 64

2 588.0 (M + 1) 65

2 602.0 (M + 1) 66

2 664.0 (M + 1) 67

4 636.0 (M + 1) 68

4 602.0 (M + 1) 69

4 588.0 (M + 1) 70

4 616.0 (M + 1) 71

2 616.0 (M + 1) 72

2 652.0 (M + 1) 73

2 658.2 (M + 1) 74

2 546.3 (M + 1) 75

2 589.2 (M + 1) 76

2 603.3 (M + 1) 77

2 635.3 (M + 1) 78

2 611.2 (M + 1) 79

2 597.3 (M + 1) 80

2 612.3 (M + 1) 81

2 613.3 (M + 1) 82

2 609.3 (M + 1) 83

2 574.0 (M + 1)

Examples 84-86

[1206] The compounds of Examples 84-85 are prepared from an appropriateA domain piperazine by following a substantially similar couplingprocedure as described in Procedures 1-5.

Coupling Example Z Procedure MS 84

2 584.2 (M + H) 85

2 586.1 (M + H)

Example 86

[1207] Example 86 is prepared by following a substantially similarcoupling procedure as described in Procedure 2.

Examples 87-100

[1208] The compounds of Examples 87-100 are prepared from an appropriateA domain piperazine by following a substantially similar couplingprocedure as described in Procedures 1-5.

Coup- ling Exam- Proce- Found ple Z dure MS (ESI) 87

4 546.3 (M + H) 88

4 638.3 (M + H) 89

4 574.3 (M + H) 90

4 572.3 (M + H) 91

4 586.3 (M + H) 92

4 614.4 (M + H) 93

4 570.3 (M + H) 94

4 584.3 (M + H) 95

4 586.2 (M + H) 96

4 614.3 (M + H) 97

4 640.3 (M + H) 98

4 640.3 (M + H) 99

4 640.3 (M + H) 100

4 640.3 (M + H)

Examples 101-102

[1209] The compounds of Examples 101 and 102 are prepared from anappropriate A domain piperazine by following a substantially similarcoupling procedure as described in Procedures 1-5.

Coupling Found Example Z Procedure MS (ESI) 101

4 570.1 (M + H) 102

4 572.1 (M + H)

Examples 103-146

[1210] The compounds of Examples 103-146 are prepared from anappropriate A domain piperazine by following a substantially similarcoupling procedure as described in Procedures 1-5.

Coup- ling Exam- Proce- Found ple Z dure MS (ESI) 103

2 601.3 (M + H) 104

2 603.3 (M + H) 105

2 651.1 (M + H) 106

2 595.2 (M + H) 107

2 601.1 (M + H) 108

2 609.3 (M + H) 109

2 668.3 (M + H) 110

2 625.3 (M + H) 111

2 609.3 (M + H) 112

2 615.2 (M + H) 113

2 629.2 (M + H) 114

2 587.2 (M + H) 115

2 619.2 (M + H) 116

2 675.3 (M + H) 117

2 623.3 (M + H) 118

2 596.2 (M + H) 119

2 585.3 (M + H) 120

2 610.2 (M + H) 121

1 624.2 (M + H) 122

1 652.3 (M + H) 123

1 692.3 (M + H) 124

2 664.3 (M + H) 125

2 650.3 (M + H) 126

2 652.3 (M + H) 127

2 666.3 (M + H) 128

2 680.3 (M + H) 129

2 574.3 (M + H) 130

2 616.3 (M + H) 131

2 632.3 (M + H) 132

2 660.3 (M + H) 133

2 674.3 (M + H) 134

2 688.4 (M + H) 135

2 588.3 (M + H) 136

2 602.3 (M + H) 137

1 622.2 (M + H) 138

2 610.0 (M + H) 139

2 638.0 (M + H) 140

2 624.2 (M + H) 141

2 658.2 (M + H) 142

2 672.0 (M + H) 143

2 625.2 (M + H) 144

2 614.2 (M + H) 145

2 615.2 (M + H) 146

2 584.0 (M + H)

Examples 147-148

[1211] The compounds of Examples 150-151 are prepared from anappropriate A domain piperazine by following a substantially similarcoupling procedure as described in Procedures 1-5.

Coupling Found Example Z Procedure MS (ESI) 147

5 600.2 (M + H) 148

5 —

Example 149-150N-(1-(4-Chloro-benzyl)-2-{4-[2-(2-isobuyl-2H-tetrazol-5-yl)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-2-(2,3-dihydro-1H-isoindol-1-yl)-acetamideandN-(1-(4-Chloro-benzyl)-2-{4-[2-(1-isobutyl-1H-tetrazol-5-yl)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-2-(2,3-dihydro-1H-isoindol-1-yl)-acetamide

[1212]

[1213] The mixture of4-[2-(2-isobutyl-2H-tetrazol-5-yl)-phenyl]piperazine and4-[2-(1-isobutyl-1H-tetrazol-5-yl)-phenyl]piperazine (60:40 by NMRfavoring the 2H substituted tetrazole, 230 mg, 0.8 mmol, 1.0 eq) wascoupled in a similar manner as described in coupling procedure 2. Theregioisomers were separated using silica gel chromatography. Theseparated compounds were deprotected using TFA followed by purificationand HCl salt formation.

[1214] 2H substituted tetrazole: HRMS (ES+) calculated for C₃₄H₄₀N₈O₂Cl:627.2963. Found: 627.2946.

[1215] 1H substituted tetrazole: HREMS (ES+) calculated forC₃₄H₄₀N₈O₂Cl: 627.2963. Found: 627.2961.

Example 1516-Hydroxy-1,1-dimethyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid{1-(4-chloro-benzyl)-2-oxo-2-[4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazin-1-yl]-ethyl}-amide

[1216]

[1217] 1-Boc-4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazine wasdeprotected and coupled to Boc-D-p-Cl-Phe-OH in a manner similar tocoupling procedure 1. The coupled product was deprotected and preparedas the chloride salt. To a solution of the chloride salt (1.16 g, 2.52mmol),6-hydroxy-1,1-dimethyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid(714 mg), DIEA (1.75 mL), HOBt (408 mg), and DMAP (62 mg) in 2.52 mL ofCH₂Cl₂ was added EDC (579 mg). After stirring overnight, the mixture wasextracted with EtOAc, washed with water, saturated bicarbonate andbrine, and then dried over Na₂SO₄, filtered and evaporated to dryness.The mixture was chromatographed with 5% MeOH/EtOAc. The diastereomerswere separated on a waters symmetry C18 column 80:20 to 50:50 water(0.05% TFA) acetonitrile over 40 minutes detecting at 230 nm. LRMS(ESI+): 628.3 (M+1).

Example 152 1-(D-TIC-4-Cl-D-Phe)-4-(2-methanesulfonatephenyl)piperazine

[1218]

[1219] To a solution of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-hydroxy-phenyl)piperazine (150 mg, 0.242mmol) and Et₃N (50 mL, 0.36 mmol) in 6 mL of CH₂Cl₂ cooled to 0° C. wasadded methanesulfonyl chloride (19 microliters, 0.24 mmol). Afterstirring for 2 hours, the reaction was quenched with saturated sodiumbicarbonate and extracted with CH₂Cl₂. The combined organic solutionswere washed with 1 M HCl, saturated sodium bicarbonate, brine, dried(Na₂SO₄), filtered, and concentrated. Without further purification, theproduct was deprotected with TFA by following the procedure as describedin Coupling Procedure 1, Step 4. HRMS (ESI+) calculated forC₃₀H₃₄ClN₄O₅S: 597.1938. Found: 597.1954 (M+H).

Example 153 1-(D-TIC-4-Cl-D-Phe)-4-(2-aminophenyl)piperazine

[1220]

[1221] A solution of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-nitro-phenyl)piperazine (260 mg, 0.4mmol), PtO₂ (70 mg) in 30 mL of isopropanol was shaken in a Parrhydrogenation apparatus under 45 psi of H₂ for about 1 hour. Thesolution was filtered through celite and concentrated to yield about 263mg (0.4 mmol, 100%) of the amine which was used without furtherpurification. The amine was deprotected with TFA by following theprocedure as described in Coupling Procedure 1, Step 4. HRMS (ESI+)calculated for C₂₉H₃₃ClN₅O₂: 518.2323. Found: 518.2338 (M+H).

Example 154 1-(D-TIC-4-Cl-D-Phe)-4-(2-sulfonamide)piperazine

[1222]

[1223] To a solution of1-(N-Boc-D-TIC-4-Cl-D-Phe)-4-(2-aminophenyl)piperazine (120 mg, 0.19mmol) and Et₃N (27 microliters, 0.19 mmol) in 6 mL of CH₂Cl₂ cooled to0° C. was added methanesulfonyl chloride (15 microliters, 0.19 mmol).After stirring for 2 hours, the reaction was quenched with saturatedsodium bicarbonate and extracted with CH₂Cl₂. The combined organicsolutions were washed with 1 M HCl, saturated sodium bicarbonate andbrine, and then dried (Na₂SO₄), filtered and concentrated. Withoutfurther purification, the product was deprotected with TFA by followingthe procedure as described in Coupling Procedure 1, Step 4. HRMS (ESI+)calculated for C₃₀H₃₅ClN₅O₄S: 596.2098. Found: 596.2104 (M+H).

Example 155N-[2-(4-{3-D-(4-chloro-phenyl)-2-D-[(1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-propionyl}-piperazin-1-yl]-methanesulfonamidetrihydrochloride

[1224]

[1225] Step 1: 1-(2-Nitrophenyl)piperazine (3.13 g, 15.1 mmol) wascoupled with Boc-D-4-chloro-phenyl alanine (4.52 g, 15.1 mmoles) in thepresence of EDC/HOBT. The crude product was chromatographed on silicagel (EtOAc/hexane 1:1) to give yellow solids (6.88 g). Mass: MH⁺ 489

[1226] Step 2:{1-4-Chloro-phenyl)-2-[4-2-nitro-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester made above (6.88 g, 14.1 mmol) was mixed with 4 MHCl in dioxane (230 ml) and stirred at r.t. for about an hour and thenconcentrated to give yellow solids (5.1 g). Mass: MH⁺ 389

[1227] Step 3:2-Amino-3-(4-chloro-phenyl)-1-[4-(2-nitro-phenyl)-piperazin-1-yl]-propan-1-onehydrochloride made above (2.5 g, 5.88 mmol) and NaOAc (1.7 g, 20.7 mmol)were dissolved in MeOH (175 ml) and cooled in an ice-water bath. Thealdehyde from Preparation 6C (2.02 g, 7.7 mmol) was added and stirredfor several minutes and then NaBH₃CN (0.48 g, 7.6 mmol) was added. Themixture was stirred at r.t. overnight. More NaOAc (0.57 g, 7.0 mmol),the aldehyde (0.67 g, 2.6 mmol) and NaBH₃CN (0.16 g, 2.5 mmol) wereadded with bath in place. The mixture was stirred at r.t. for about 4hours, and then stripped to dryness. 1 M HCl and EtOAc were addedfollowed by washing with NaHCO₃ and brine, and then dried over Na₂SO₄.Removal of solvent gave a residue, which was chromatographed on silicagel (2% MeOH/CH₂Cl₂) to give yellow solids (2.53 g). Mass: MH⁺ 634

[1228] Step 4:3-({1-(4-Chloro-benzyl)-2-[4-(2-nitro-phenyl)-piperazin-1-yl]-2-oxo-ethylamino}-methyl)-3,4-1H-isoquinoline-2-carboxylicacid tert-butyl ester made above (2.5 g, 3.94 mmol) was dissolved inCH₂Cl₂ (10 ml) and cooled to 0° C. TEA (0.4 g, 4.0 mmoles) and Bocanhydride (0.86 g, 3.94 mmol) dissolved in CH₂Cl₂ (10 ml) was added tothe mixture dropwise. Additional TEA (0.4 g, 4.0 mmol) was added and themixture was stirred for about 1.5 hours. The mixture was concentrated toremove Et₃N and CH₂Cl₂ was added. The mixture was stirred over theweekend. Additional DMAP (0.096 g, 0.79 mmol) and TEA (0.4 g, 4.0 mmol)were added and the mixture was stirred for about 5 hours. The mixturewas stripped to dryness and chromatographed with ethyl acetate/hexane(2:8) to afford about 1.06 g of product. Mass: MH⁺ 734

[1229] Step 5:3-[tert-Butoxycarbonyl-{1-(4-chloro-benzyl)-2-[4-2-nitro-phenyl)-piperazin-1-yl]-2-oxo-ethyl}-amino)-methyl]-3,4-dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester made above (0.50 g, 0.68 mmol) was dissolved inisopropyl alcohol (100 ml) and Pt₂O (0.13 g, 0.59 mmol) was added. Thehydrogenation was carried out on Parr shaker at 45 psi for about an hourat r.t. The mixture was filtered, stripped to dryness to give a whitesolid (0.46 g). Mass: MH⁺ 704

[1230] Step 6:3-({[2-[4-(2-Amino-phenyl)-piperazin-1-yl]-(4-chloro-benzyl)-2-oxo-ethyl]-tert-butoxycarbonyl-amino}-methyl)-3,4dihydro-1H-isoquinoline-2-carboxylicacid tert-butyl ester made above (0.46 g, 0.65 mmol) was dissolved inCH₂Cl₂ (10 ml). The mixture was cooled with an ice bath under nitrogen,and then TEA (0.13 g, 1.31 mmoles) was added followed by slow additionof MsCl (0.075 g, 0.65 mmol) in CH₂Cl₂ (1 ml). After about 30 minutes,an additional amount of MsCl (0.025 g, 0.22 mmol) was added. The mixturewas cooled, diluted with ethyl acetate, extracted with saturated Na₂CO₃,washed with brine, dried and evaporated in vacuo. The material waschromatographed on ion exchange chromatography (0.35 g). Mass: MH⁺ 782

[1231] Step 7:3-[tert-Butoxycarbonyl-{1-(4-chloro-benzyl)-2-[4-(2-methanesulfonylamino-phenyl)-piperazin-1-yl]2-oxo-ethyl}-amino)-methyl]-3,4-dihydro-1H-isoquinoline-2-carboxylicacid t-butyl ester made above (0.35 g, 0.65 mmol) was stirred with 4MHCl in dioxane (30 ml) at r.t. for about an hour. The mixture wasstripped to dryness and saturated sodium bicarbonate was added. Themixture was then extracted with ethyl acetate, washed with brine anddried. The material was chromatographed on silica gel using 5%MeOH/CH₂Cl₂. The residue was dissolved in methanol (40 ml) and 2M HCl inether (3 ml) was added, which was then stripped to dryness affordingabout 0.23 g of the final compound. Calculated exact mass: 582.2305.Found exact mass: 582.2286

Example 1562-(4-{3-D-(4-chloro-phenyl)-2-D-[(1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-propionyl}-piperazin-1-yl)-benzenesulfonamidetrihydrochloride

[1232]

[1233] The aldehyde from Preparation 6C was reacted with2-{4-[2-Amino-3-D-(4-chloro-phenyl)-propionyl]-piperazin-1-yl}-benzenesulfonamidehydrochloride by following the procedure described in Example 158, Step3 and then Step 7. Deprotection of Boc group in the presence of 4MHCl/dioxane gave the title compound. Exact mass calculated: 568.2419;Found: 568.2158.

Example 1573-(4-Chloro-phenyl)-2-[methyl-(1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-1-[4-(2-pyrrolidin-1-ylmethyl-phenyl)-piperazin-1-yl]-propan-1-onetetrahydrochloride

[1234]

[1235] Step A: A 4M solution of HCl in dioxane (20 mL) was added to asolution of 4-(2-Pyrrolidin-1-ylmethyl-phenyl)-piperazine-1-carboxylicacid t-butyl ester (2.01 gm, 5.82 mmol). The solution was stirred atr.t. overnight under a nitrogen and then concentrated to remove dioxane.Diethyl ether was added and the solution was concentrated (2×). Diethylether was added and the product was isolated by suction filtration andthen washed with diethyl ether. Vacuum drying at 50° C. overnight gave1-(2-Pyrrolidin-1-ylmethyl-phenyl)-piperazine 2HCl (1.62 g, 87.6%). MS(m/z, ES+): 246.1.

[1236] Step B: Lithium2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-methyl-amino]-3-(4-chloro-phenyl)-propionate(0.59 gm, 1.27 mmol), the compound of Step A (0.27 gm, 0.85 mmol), EDC(0.24 gm, 1.27 mmol) and HOBt (0.17 gm, 1.27 mmol) were combined anddissolved in anhydrous DMF (5 mL). DIPEA was added (440 microliter, 2.54mmol), and the reaction was stirred under nitrogen overnight at roomtemperature. The reaction was concentrated and reconstituted in CH₂Cl₂and then diluted with NaHCO₃. After separation of the organic phase, theaqueous layer was extracted with CH₂Cl₂ (2×). The combined organics weredried (Na₂SO₄), filtered and concentrated. Chromatography (EtOAc to 5%MeOH/EtOAc) gave about 100 mg of3-[({1-(4-Chloro-benzyl)-2-oxo-2-[4-(2-pyrrolidin-1-ylmethyl-phenyl)-piperazin-1-yl]-ethyl}-methyl-amino)-methyl]-3,4-dihydro-1H-isoquinoline-2-carboxylicacid t-butyl ester. MS (m/z, ES+): 686.4.

[1237] Step C: The material from Step B was taken up in a 4M solution ofHCl in dioxane (30 mL). The reaction was stirred at r.t. overnight undernitrogen. The mixture was concentrated to remove dioxane, and theresulting film was triturated with diethyl ether and then concentrated(2×). Trituration with diethyl ether, isolation by suction filtration,and drying at r.t. under vacuum gave about 0.103 g of the final compoundas yellow solids (97%). MS (m/z, ES+): 586.3.

Example 1583-(4-Chloro-phenyl)-2-[(2-methoxy-ethyl)-(1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-1-[4-(2-pyrrolidin-1-ylmethyl-phenyl)-piperazin-1-yl]-propan-1-onetetrahydrochloride

[1238]

[1239] 4-(2-Pyrrolidin-1-ylmethyl-phenyl)-piperazine-1-carboxylic acidt-butyl ester was deprotected and then the resulting amine hydrochloride(0.10 gm, 0.30 mmol) was coupled with lithium2-[(2-tert-butoxycarbonyl-1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-(2-methoxy-ethyl)-amino]-3-(4-chloro-phenyl)-propionate(0.23 gm, 0.45 mmol). The mixture was chromatographed to obtain thecrude coupled product [MS (m/z, ES+): 730.4], which was deprotected togive about 0.068 g of the final compound as brown solids. MS (m/z, ES+):630.3

Example 159(R)-N-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(2-([1,2,4]triazol-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-2-(2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetamide(isomer 1)

[1240]

[1241] To a solution of2-amino-3-(4-chloro-phenyl)-1-[4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazin-1-yl]-propan-1-onetrifluoroacetyl carboxylate salt(0.30 g, 0.55 mmol),(2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid (0.12 g, 0.55mmol), HATU (0.21 g, 0.55 mmol) in DCM was added DIPEA (0.19 mL, 1.13mmol). After about 3 hours, the solution was purified by silica gelchromatography (eluent: 3% 2.0M NH₃ in MeOH/DCM). The purified fractionswere combined and concentrated under reduced pressure to give the finalcompound as white foam (0.06 g, 18%). ES MS 626.3 (M+H)

Example 160(R)-N-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(2-([1,2,4]triazol-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-2-(2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetamidedihydrochloride salt (isomer 1)

[1242]

[1243] Into a flask containing(R)-N-{1-(4-chlorobenzyl)-2-oxo-2-[4-(2-([1,2,4]triazol-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-2-(2-isopropyl-2,3-dihydro-1H-isoindol-1-yl)-acetamide(Example 162) was added 1.0 N HCl (5 mL). After about an hour, thesolution was solidified at −78° C., and the solid lyophilized to giveabout 0.06 g of the final compound as tan solids. ES MS 626.3 (M+H)

Example 1612-(2-Butyl-2,3-dihydro-1H-isoindol-1-yl)-N-{1-(4-chlorobenzyl)-2-oxo-2-[4-((2-[1,2,4]triazol-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-acetamide(isomer 1)

[1244]

[1245] To a solution prepared from Preparation 4AB (0.30 g., 0.45 mmol),(2-butyl-2,3-dihydro-1H-isoindol-1-yl)-acetic acid (Preparation 8C)(0.10 g, 0.45 mmol), HATU (0.17 g., 0.45 mmol) in DCM (5.1 mL) was addedDIPEA (0.16 mL, 0.91 mmol). After about 3 hours, the solution waspurified by silica gel chromatography (eluent: 2-4% 2.0M NH₃ inMeOH)/DCM). The purified fractions were combined and concentrated underreduced pressure to give about 0.07 g of the final compound as off-whitefoam (26%). ES MS 640.3 (M+H)

Example 1622-(2-Butyl-2,3-dihydro-1H-isoindol-1-yl)-N-{1-(4-chlorobenzyl)-2-oxo-2-[4-((2-[1,2,4]triazol-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-acetamidedihydrochloride salt (isomer 1)

[1246]

[1247] To a flask containing2-(2-butyl-2,3-dihydro-1H-isoindol-1-yl)-N-{1-(4-chlorobenzyl)-2-oxo-2-[4-((2-[1,2,4]triazol-1-yl)methylphenyl)-piperazin-1-yl]-ethyl}-acetamide(Example 166)(0.07 g, 0.11 mmol) was added 1.0 N HCl (5 mL). After aboutan hour, the solution was solidified at −78° C., and the solidlyophilized to give about 0.06 g of the final compound as green solids.ES MS 640.3 (M+H)

Examples 163-166

[1248] The Examples 163 to 166 were prepared as follows. The mixture of4AB-2TFA salts or 4AB-HCl salts (Preparation 4AB) (1.0 eq.),N-Boc-substituted-D-Tic-OH or N-Boc-substituted-DL-Tic-OH (1.0 eq.),HATU (1.0 eq.) and DIEA (5.0-10.0 eq.) in DCM was stirred at r.t.overnight. The mixture was partitioned between water and CH₂Cl₂. Theaqueous layer was extracted with CH₂Cl₂ (2×). The combined organicsolution was dried over MgSO₄, filtered and concentrated in vacuo. Themixture purified by silica gel column using 10% MeOH in EtOAc to giveN-Boc product.

[1249] The N-Boc product was mixed with 5 mL of saturated HCl in EtOAcand stirred at r.t. overnight. Diethylether was added, and the resultingwhite solid was filtered and washed with ether (3×) to give the finalcompound as HCl salt.

Example 163 3-Methyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid{1-(4-chloro-benzyl)-2-oxo-2-[4-(2-[1,2,4]triazol-yl-methyl-phenyl)-piperazin-1-yl]-ethyl}-amide,HCl Salt

[1250]

Example 164 7-Fluoro-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid{1-(4-chloro-benzyl)-2-oxo-2-[4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazin-1-yl]-ethyl}-amide,HCl Salt

[1251]

Example 1657-Trifluoromethyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid{1-(4-chloro-benzyl)-2-oxo-2-[4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazin-1-yl]-ethyl}-amide,HCl Salt

[1252]

Example 166 3-Methyl-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid{1-(4-chloro-benzyl)-2-oxo-2-[4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazin-1-yl]ethyl}-amide,HCl Salt

[1253]

Example 167 1,2,3,4-Tetrahydro-isoquinoline-3-carboxylic acid[2-{4-[2-(isobutyl-methanesulfonyl-amino)-phenyl]-piperazin-1-yl}-1-(4-methoxy-benzyl)-2-oxo-ethyl]-amide,2HCl Salt (isomer 2)

[1254]

[1255] The above compound is prepared from the A domain 98A (Preparation98A) and the BC domain from Preparation 11BC following a proceduresubstantially similar to Coupling Procedure 2. LRMS (ESI+): 648.3 (M+H)

Example 1682-(2,3-Dihydro-1H-isoindol-1-yl)-N-(1-(4-fluoro-benzyl)-2-{4-[2-(isobutyl-methanesulfonyl-amino)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-acetamide,HCl Salt

[1256]

[1257] The above compound was prepared by following substantiallysimilar procedure as described in Example 167. MS M+1 636.3 (90%).

Example 1693-(4-Chloro-phenyl)-1-[4-(2-dimethylaminomethyl-phenyl)-piperazin-1-yl]-2-[methyl-(1,2,3,4-tetrahydro-isoquinolin-3-ylmethyl)-amino]-propan-1-onetri-hydrochloride salt

[1258]

[1259] The Boc protected compound of Example 90 (0.19 g, 0.29 mmol) wasdissolved in MeOH and stirred under N₂ at room temperature. NaOAc (0.12g, 1.5 mmol) was added to the mixture followed by aqueous HCHO (0.11 ml,1.5 mmol). The mixture was stirred at r.t. for about 30 minutes. NaBH₃CN(0.06 g, 88 mmol) in MeOH (2 ml) was added dropwise at 0° C. The mixturewas stirred at r.t. for about an hour. The mixture was concentrated,taken up in EtOAc and washed with dilute NaHCO₃ and brine. The mixturewas dried over Na₂SO₄ and solvent was evaporated. The resulting residuewas purified by flash chromatography (silica gel, 6% 2M NH₃/MeOH/CH₂Cl₂)to give about 0.3 g of Boc protected amine compound (2) as white solids.Mass: MH⁺ 660

[1260] To the compound obtained above (0.18 g) was added 4M HCl/dioxane(15 ml) and the mixture was stirred at r.t. for about 20 minutes. Themixture was stripped to dryness and triturated with Et₂O to afford about0.24 g of the final compound as white solids (92%). LC-MS: MH⁺ 560;Exact mass calculated: 560.3156; Pound: 560.3170.

Example 170 1-(D-TIC-4-Cl-D-Phe)-4-[(2-(1-S hydroxyethyl)phenyl]piperazine

[1261]

[1262] The A domain piperazine of Preparation 11A was coupled toBoc-D-TIC-4-Cl-D-Phe-OH in a manner substantially similar to thatdescribe in Coupling Procedure 2. To a solution of the protected product(100 mg, 0.131 mmol) in 2 mL CH₂Cl₂ was added 1 drop of H₂O, and 1 mL ofTFA. After stirring at r.t. for about 3 hours, the solution isazeotroped from heptane (3×). To a solution of the residue in THF at 0°C. was added 1 mL of HF-pyr. After stirring overnight, the solution wasdiluted with CH₂Cl₂, washed with saturated sodium bicarbonate (2×) andbrine, and then dried (Na₂SO₄), filtered and concentrated. Afterpurification by flash chromatography (10 g SiO₂, linear gradient 0-10%methanol/CH₂Cl₂, 30 mL/minute over 30 minutes), the product wasdissolved in CH₂Cl₂ and precipitated with 1 M HCl in Et₂O to affordabout 63 mg (0.11 mmol, 82%) of the final compound. HRMS (electrospray)calculated for C₃₁H₃₆ClN₄O₃: 547.2476. Found: 547.2485 (M+H).

Example 171 1-(D-TIC-4-Cl-D-Phe)-4-[(2-(1-R hydroxyethyl)phenyl]piperazine

[1263]

[1264] The A domain piperazine of Preparation 12A was coupled toBoc-D-TIC-4-Cl-D-Phe-OH and deprotected in a manner substantiallysimilar to that describe in Example 171 above. HRMS (electrospray)calculated for C₃₁H₃₆ClN₄O₃: 547.2476. Found: 547.2480 (M+H).

[1265] The following Examples 172-174 are prepared from an appropriatelysubstituted A domain piperazine by following a substantially similarcoupling procedure as described in Coupling Procedures 1.

Example 172 Isoquinoline-3-carboxylic acid{1-(4-chloro-benzyl)-2-oxo-2-[4-(2-[1,2,4]triazol-1-ylmethyl-phenyl)-piperazin-1-yl]-ethyl}-amide

[1266]

[1267] The above compound is prepared by following a CouplingProcedure 1. Found MS (ESI) 580.2 (M+H)

Example 173 Isoquinoline-3-carboxylic acid(1-(4-chloro-benzyl)-2-oxo-2-{4-[2-(propionylamino-methyl)-phenyl]-piperazin-1-yl}-ethyl)-amide

[1268]

[1269] The above compound is prepared by following a CouplingProcedure 1. Found MS (ESI) 584.3 (M+H)

Example 174 Isoquinoline-3-carboxylic acid(1-(4-chloro-benzyl)-2-{4-[2-(N-isobutyl-hydrazino)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-amide

[1270]

[1271] The above compound was prepared by following a CouplingProcedure 1. Found MS (ESI) 648.0 (M+H)

[1272] Preparation of Novel C-Domain Pieces

[1273] Heck Coupling:

Preparation PP1

[1274] Synthesis of Compound (2a) by a Heck Coupling of2-bromobenzaldehyde (1a) with methyl acrylate (Pd(OAc)₂/PPh₃ as thecatalyst): A mixture of 2-bromobenzaldehye (1a) (24.5 g, 132 mmol),methyl acrylate (17.9 mL, 199 mmol), Pd(OAc)₂ (590 mg, 2.65 mmol, 2 mol%), PPh₃ (1.39 g, 5.30 mmol, 4 mol %) and Et₃N (46 mL, 331 mmol) wasstirred at 80° C. for 15 h. Large amount of yellow solid was formedafter the reaction was done. The mixture was cooled to rt, concentrated,and mixed with H₂O (200 mL). The organic solid was collected byfiltration, and then applied to a plug of silica gel (25 g)(EtOAc/hexane 1:1) to give a dark yellow solid. The solid was purifiedby crystallization (100 mL EtOAc bottom layer, 120 mL hexane top layer)to provide 17.57 g (70%) (100% pure by NMR) of the first crop and 5.23 g(21%) (95% by NMR) of the second crop of 2a.

Preparation PP2

[1275] Synthesis of Compound (2a) by a Heck Coupling of2-bromobenzaldehyde (1a) with Methyl Acrylate (R═H)(Pd(OAc)₂/P(O-Tolyl)₃ as the catalyst): The compound 1a (9.998 g, 54.04mmol) was dissolved in toluene (20 mL) at r.t. Methylacrylate (5.996 g,69.65 mmol, 1.29 eq.), NEt₃ (15 mL), Pd(OAc)₂ and P(O-Tolyl)₃ weresuccessively added and the mixture was stirred under reflux. After 2hours, the reaction mixture was allowed to cool to r.t. Then theprecipitated yellow catalyst was removed by filtration. The catalyst wasrinsed with toluene (2×10 mL) and the filtrates were concentrated todryness under reduced pressure. The residual oil was dried under vacuumover the weekend to give a crude solid (11.449 g). The solid wastaken-up with isopropanol (25 mL) and stirred overnight at r.t. Then,the precipitate was filtered and rinsed with isopropanol (5 mL). The wetcake (8.240 g) was dried overnight at RT affording the highly pure2-carboxaldehyde-methyl-cinnamate with 74% yield (7.627 g, 40.1 mmol).

Preparation PP3

[1276] Heck Coupling of 1b and methyl acrylate to form 2b (R=5-OMe): Amixture of 2-bromo-5-methoxybenzaldehyde (1b) (4.5 g, 20.9 mmol,Aldrich), methyl acrylate (2.7 g, 1.5 eq, 2.83 mL), Et₃N (7.4 g, 3.5 eq,10.2 mL), Pd(OAc)₂ (93 mg, 0.02 eq), and P(O-Tol)₃ was stirred andheated to 80° C. over 2-3 days. The reaction mixture was cooled to r.t.,partitioned between EtOAc (50 mL) and brine (50 mL). The aqueous wasextracted with EtOAc (2×50 mL). The combined organic was washed withbrine (1×50 mL), dried over MgSO₄, filtered, concentrated to yield ayellow brown oil (5.01 g, 109%). This crude oil was purified in a hotsolvent Hex/EtOAc (80 mL/15 mL) to yield 2b as a pale yellow solid (3.5g, 76%).

Preparation PP4

[1277] Heck Coupling of 1c and Methyl Acrylate to Form 2c (R=4,5-OMe):To a solution of 1c (906 mg, 3.70 mmol) in toluene (2 mL) was addedPd(OAc)₂ (17 mg, 0.074 mmol, 2 mol %), P(O-Tolyl)₃ (45 mg, 0.148 mmol, 4mol %), methyl acrylate (0.5 mL, 5.55 mmol) and Et₃N (1.5 mL, 11.1mmol). The mixture was stirred at 80° C. for 21 h, cooled to rt, andmixed with H₂O (40 mL). The organic compounds were extracted with EtOAc(50 mL), washed with brine (40 mL), dried (Na₂SO₄), and concentrated.The residue was purified by flash chromatography to provide 466 mg (47%)of recovered 1c followed by 450 mg (49%) of 2c (4,5-Ome).

Preparation PP5

[1278] Heck Coupling of 1d and Methyl Acrylate to Form 2d (R=5-NO₂): Theprocedure is same as that of 2c, yielding 82% of 2d after purification.

Preparation PP6

[1279] Reductive Amination

[1280] Reductive amination of (2a) with benzyl amine to form isoindoline(10a). To a solution of 2a (11.27 g, 59.2 mmol) in ClCH₂CH₂Cl (60 mL)was added BnNH₂ (6.47 mL, 59.2 mmol), followed by HOAc (5.1 mL, 89mmol). The mixture was stirred at rt for 1 h. NaCNBH₃ (5.58 g, 88.8mmol) and MeOH (30 mL) were then added to the above solution. Theresulting mixture was stirred at rt for another 2 h and quenched withsat. NaHCO₃ solution (150 mL). The mixture was extracted with EtOAc(2×100 mL) and the combined organic layers were washed with brine (150mL), dried (Na₂SO₄), and concentrated to provide 15.3 g of crude productof 10a which was carried out for the next hydrogenolysis reaction.

Preparation PP7

[1281]

[1282] One-pot process from 2-carboxaldehyde-methyl-cinnamate to targetcyclized isoindoline product using NaBH₃CN.2-carboxaldehyde-methyl-cinnamate 2a (3.254 g, 17.1 mmol) was dissolvedin a 1:1 MeOH:PhCH₃ mixture (20 mL) at r.t. R-(+)-phenethylamine (2.073g, 17.1 mmol) was added and the solution was heated under reflux for 2hours. HPLC in process control indicated that the imine formation wascompleted. Then, AcOH (2.055 g, 34.2 mmol) and NaBH₃CN (2.15 g, 34.2mmol) were successively added at RT, the reaction mixture being cooledwith a water-bath. The reaction mixture was post-agitated overnight.Water (10 mL), MeOH (20 mL) and 37% HCl (2.8 mL) were successively addedand the organic layer was extracted. The aqueous layer was washed withPhCH₃ (10 mL). Then, the aqueous layer was made basic with 5N NaOH (20mL) and MeOH was concentrated to partly remove MeOH. Extraction withEtOAc (2×25 mL) was performed. The combined organic layers were driedover MgSO4, filtered and rinsed with EtOAc (10 mL). The filtrates wereconcentrated under reduced pressure and the residual oil was dried undervacuum overnight at RT to afford the target cyclized isoindoline product10b with 92% yield (4.642 g, 15.7 mmol). HPLC % area indicated that the2 diastereomers were produced in a 55:45 ratio. ¹H NMR confirmed thisresult by integration of the methyl group of the phenethyl substituent.Note: The Heck or Heck-type coupling was performed in toluene with aslight excess of methylacrylate which was removed by distillation beforethe MeOH and the R-(+)-phenethylamine addition.

[1283] Reductive amination of (2a) with t-butyl carbamate to form (11a):To a solution of aldehyde 2a (238 mg, 1.25 mmol) in CH₃CN (8 mL) wasadded t-butyl carbamate (439 mg, 3.75 mmol), followed by triethylsilane(0.6 mL, 3.75 mmol) and TFA (0.19 mL, 2.5 mmol). The mixture was stirredat rt overnight, quenched with sat. NaHCO₃ solution (20 mL) andextracted with EtOAc (2×30 mL). The combined organic layers were washedwith brine (30 mL), dried (Na₂SO₄) and concentrated. The residue waspurified by flash chromatography (hexane/EtOAc 3:1) to provide 317 mg(87%) of 11a.

[1284] Reductive amination of 2b with t-butyl carbamate to form 11b: Amixture of aldehyde 2b (600 mg, 2.72 mmol) Et₃SiH (955 mg, 3 eq, 1.31mL), TFA (620 mg, 2 eq, 420 uL), t-butyl carbamate (980 mg, 3 eq) inacetonitrile (15 mL) was stirred at room temperature over 2 days.Removed the solvent on a Rotary evaporator and purified the cruderesidue on a flash column (100 g SiO₂, 7:1→6:1 Hex/EtOAc). Collected 307mg good desired product 11b (35%); 195 mg product contaminated withaldehyde SM (22%).

[1285] Reductive amination of (2c) with t-butyl carbamate to form (11c):To a solution of aldehyde 2c (411 mg, 1.64 mmol) in CH₃CN (10 mL) wasadded t-butyl carbamate (580 mg, 4.93 mmol), followed by triethylsilane(0.8 mL, 4.93 mmol) and TFA (0.25 mL, 3.28 mmol). The mixture wasstirred at rt overnight, quenched with sat. NaHCO₃ solution (30 mL) andextracted with EtOAc (2×30 mL). The combined organic layers were washedwith brine (30 mL), dried (Na₂SO₄) and concentrated. The residue waspurified by flash chromatography (hexane/EtOAc 3:1, hexane/EtOAc 1:1) toprovide 535 mg (93%) of 11c.

[1286] To a solution of 2d (1.02 g, 4.34 mg) in CH₂Cl₂/CH₃CN (1:1 24 mL)was added BocNH₂ (1.5 g, 13.02 mmol), Et₃SiH (2.1 mL, 13.02 mmol), andTFA (0.67 mL, 8,67 mmol). The mixture was stirred at rt for 7 h. Aprecipitate was formed during the reaction. The reaction mixture wasquenched with sat. NaHCO₃ solution (30 mL), and diluted with CH₂Cl₂ (40mL). The organic layer was washed with brine (30 mL), dried (Na₂SO₄),and concentrated. The residue was purified by flash chromatography(hexane/EtOAc 3:1, then CH₂Cl₂/EtOAc 10:1) to provide 2.08 g yellowsolid which still containing BocNH₂. The product is not the desiredBoc-carbamate 14c. LC-MS result showed that the product is the Schiffbase intermediate.

[1287] To the above product (420 mg) in CH₂Cl₂ (10 mL) was added Et₃SiH(1 mL) and TFA (0.4 mL). The mixture was stirred at rt for 1 h and smallamount of sample was taken for NMR. NMR analysis demonstrated that thestarting material was consumed and the product was 14c. TFA (0.7 mL) wasthen added to the above mixture and the resultant solution was stirredat rt for another 5 h and concentrated. The residue was dissolved inEtOAc (20 mL) and washed with H₂O (10 mL). The aqueous layer wasbasified with sat. NaHCO₃ (30 mL) and the organic compounds wereextracted with CH₂Cl₂ (2×25 mL). The combined organic layers were washedwith brine (20 mL), dried (Na₂SO₄) and concentrated to provide 218 mg ofthe cyclized compound 14c.

[1288] Condensation of 2a with alpha-methylbenzylamine to Form Imine 9.2-carboxaldehyde-methyl-cinnamate 2a (0.897 g, 4.72 mmol) was dissolvedin MeOH (10 mL) at r.t. R-(+)-phenethylamine (0.577 g, 4.76 mmol) wasadded and the solution was heated under reflux for 2 hours. HPLC inprocess control indicated that the imine formation was completed. Thesolvent was stripped on a rotary evaporator and the resulting oil wasdried at RT under vacuum overnight. The Schiff base 9 was obtainedalmost quantitatively (1.412 g, 4.81 mmol).

Preparation PP13

[1289] Michael Addition:

[1290] The compound of alpha-methyl benzylamine was applied as theauxiliary. As shown above, the one-pot reaction of aldehyde 2a andalpha-methyl benzylamine gave 90% of 10b with a ratio of 1.2:1.

[1291] Step-Wise Reduction, Amination and Cyclization:

[1292] Condensation of aldehyde 2a with alpha-methylbenzylamine inacetonitrile, methanol, methanol/toluene(1:1) or toluene afforded imine9 in excellent yield. Reduction of the imine was initially carried outat RT with NaCNBH₃/HOAc. As a result, a poor ee ratio (1.2:1) wasobtained, similarly to the previous described one-pot procedure. Butwhen the reaction was carried out with NaBH₄/TFA at RT, the ratio waselevated to 2:1. By lowering the reaction temperature to −78° C., theratio was increased to 5 to 6:1.

Preparation PP14

[1293] Cyclization of t-Butyl carbamate (11a): The N-Boc isoindolinemethyl ester 12 was originally synthesized from 11a via deprotection ofBoc with TFA, followed by basic workup, and protection with a Boc group.This procedure has been greatly improved by a one-step procedure.

[1294] In a 3 L 3-neck round bottom flask equipped with a nitrogeninlet, thermocouple and mechanical stirrer, a solution of 160 g (1.15moles) of K₂CO₃ in 180 mL of water was stirred at ρ.τ. Solid BOCanhydride 120 g (0.55 moles) was added in one portion forming asemi-solution. To the reaction mixture, a solution of the crude aminoester starting material, 87 g (0.46 moles) in 120 mL of THF was addedslowly at such a rate to keep the internal temperature below 35° C. Amild effervescence was observed. The reaction mixture was stirred for 18hours at ρ.τ. Analysis of a reaction aliquot via NMR (DMSO₆) indicatesthe desired product. The reaction was diluted with brine and the productextracted with EtOAc. The organic layer was dried over Na₂SO₄, filtered,and concentrated to yield a dark oil, 150.1 g, >100% yield. The crudematerial was taken on to the next step.

[1295] In a 3-L 3-neck round bottom flask equipped with a mechanicalstirrer, thermocouple, and reflux condenser, a solution of 150 g(approx. 0.46 moles) of crude N-BOC ester starting material in 750 mL ofmethanol was stirred at ρ.τ. To the solution, 750 mL of water was addedand the cloudy mixture was stirred vigorously. Solid LiOH 25 g (1.03moles) was added in small portions at such a rate to maintain theinternal temperature below 45° C. Upon completion of addition, thereaction was stirred overnight at rt becoming a dark green color. After18 hours the reaction was concentrated to yield a thick semisolid. Thecrude product was dissolved in EtOAc and washed with 1 N HCl quickly,followed by two brine washes. The organic layer was dried with Na₂SO₄,filtered and concentrated to yield 81 g of a dark green solid. Theaqueous layers were combined and back extracted with methylene chloride,dried over Na₂SO₄, filtered, and concentrated to yield 6 g of a darkgreen solid. Both solids were combined to yield 87 g of desired productconfirmed via NMR (DMSO₆).

Prearation PP17

[1296]

[1297] Synthesis of 14b: Dissolved the N-boc compound 11b (200 mg, 0.62mmol) in CH₂Cl₂ (1.0 mL). Cooled the clear light yellow solution to 0°C. Added slowly TFA (˜710 mg, 10 eq, ˜500 microliter) via a syringe.Removed the cooling bath and stirred the clear light brown solution atRT overnight. TLC (3:1 Hex/EtOAc, UV) confirmed a complete reaction.Removed the TFA on a rotavapor. Added EtOAc and concentrated again(twice). The crude residue was partitioned between EtOAc (10-15 mL) anda sat. NaHCO₃ (10-15 mL). The aqueous was extracted with EtOAc (2×10mL). The combined organic was dried over MgSO₄, filtered, andconcentrated to yield a light brown wet solid (212 mg, 138%). NMR(CD₃OD) confirmed the desired isoindoline 14b. This crude isoindolinewas used in the next protection step without purification.

Preparation PP18

[1298] Synthesis of 12b: To a mixture of the isoindoline 14b (190 mg,0.859 mmol), K₂CO₃ (189 mg, 1.5 eq) in a solvent 1:1 TFH/H₂O (1.0 mL) atRT was added BOC₂O (210 mg, 1.1 eq). The reaction mixture was stirred atRT overnight. TLC (3:1 Hex/EtOAc, UV) confirmed a complete reaction.Diluted the mixture with EtOAc (15 mL), and washed with H₂O (1×20 mL).The aqueous was extracted with EtOAc (1×20 mL). The combined organic waswashed with brine (1×20 mL), dried over MgSO₄, filtered, concentrated toyield a clear brown oil (340 mg, 123%). This crude oil was purified on aprep TLC plate (2×1,000 micron, solvent 2:1.5:0.5 CHCl₃/Hex/EtOAc) toyield 12b a clear yellow oil (190 mg, 69%). ¹H and ¹³C NMR (CDCl₃) wereobtained.

Procedure PP19

[1299] Synthesis of 12d (5-NO₂) by Boc-protection. The compound wasprepared by following the same procedure as described for 12b.

[1300] The imine 9 (1.412 g, 4.81 mmol) was dissolved in anhydrous THF(10 mL) at RT and TFA (5 mL) was added. The black solution was thencooled to −78° C. (dry ice bath) and NaBH₄ (0.893 g, 23.6 mmol, 5 eq.)was added in 2 portions over 5 minutes. Then, the reaction mixture waspost-agitated at −78° C. for 3 hours and allowed to gently warm at RTovernight. Water (20 mL), cyclohexane (10 mL) and EtOH (20 mL) weresuccessively added and the organic layer was extracted and discarded.The aqueous layer was made basic with 5N NaOH (20 mL) and extracted twotimes with a 2:1 EtOAC/PhCH₃ mixture (30 mL). The combined organiclayers were dried over MgSO4, filtered and rinsed with EtOAc (10 mL).The filtrates were concentrated under reduced pressure and the residualoil was dried under vacuum overnight at RT to afford the target cyclizedisoindoline product 10b (1.273 g, 4.31 mmol) with 91.4% yield. HPLC %area indicated that the 2 diastereomers were produced in a 84:16 ratio(de 68%). ¹H NMR confirmed this result by integration of the methylgroup of the phenethyl substituent.

Preparation PP20

[1301]

[1302] N-Boc methyl ester 11a (36.3 g, 0.125 mol) was dissolved in THF(250 mL), and the solution was cooled to about 0° C. A solution ofpotassium bis(trimethylsilyl) amide (1.24 g, 0.05 mol. Eq.) was addedslowly via a syringe under nitrogen atmosphere. The temperature wasraised about 8 degrees during the addition. The cooling bath was removedand the solution was stirred at r.t. for 30-45 min. The clear brownsolution was poured into a separation funnel containing about 100 mL ofa saturated NH₄Cl. The layers were separated. The aqueous layer wasextracted with EtOAc (2×50 mL). The combined organic was washed withbrine (1×100 mL), dried over Na₂SO₄, filtered, concentrated on a Rotaryevaporator to a clear yellow oil (37.3 g). This crude oil was purifiedon a flash column (600 g SiO₂), with a gradient solvent 6:1 Hex/EtOAc(2.1 L), 5:1 Hex/EtOAc (1.2 L), 4:1 Hex/EtOAc (1.5 L) to yield 12a as aclean yellow oil (34.5 g, 95%).

Preparation PP21

[1303]

[1304] To a solution of 11c (535 mg, 1.52 mmol) in THF (10 mL) was addedKHMDS (0.5 M in toluene, 0.1 mL, 0.05 mmol, 2 mol %). The mixture wasstirred at r.t. for 20 min, quenched with sat. NH₄Cl solution (20 mL),and diluted with EtOAc (20 mL). The organic layer was separated, washedwith brine (20 mL), dried (Na₂SO₄) and concentrated. The residue wasfiltered through a plug of silica gel (EtOAc/CH₂Cl₂ 1:10) to give 530 mg(99%) of 12c as an off white solid.

Preparation PP22

[1305] Deprotections:

[1306] Hydrogenolysis of 10a (R=Bn) to Form (14a): To a solution ofcrude 10a (15.3 g, 54.4 mmol) in MeOH (100 mL) was added Pd(OH)₂/C(Pearlman's catalyst, 1.02 g, 6 mol %) in a par-shaker bottle. Thesuspension was shaken under 30 psi H₂ pressure overnight in thepar-shaker, and filtered through a plug of celite. The filtrate wasconcentrated to provide 10.1 g of crude 14a as brown oil. (The procedureis same for the methyl benzylamine isoindoline substrate 10b).

Preparation PP23

[1307]

[1308] In a typical reaction a mixture of the isoindoline ester 12a (92mg, 0.316 mmol) in 1:1 MeOH/H₂O (2 ml) was treated with LiOH (15 mg, 2eq) at RT overnight. Diluted the mixture with CH₂Cl₂ (5 ml) and water (5ml). Adjusted the pH of the reaction mixture to 1-3 with a 10% NaHSO₄solution. Separated the layers. The aqueous was extracted with CH₂Cl₂(1×10 ml). The combined organic was dried over Na₂SO₄, filtered,concentrated to yield 16a as a pale yellow foam (76 mg, 87%). NMR(CDCl₃) showed a clean desired acid product.

[1309] It is noted that he reaction time must be more than 6 hours. Thecrude foam can be purified by slurry in warm hexane and then filter toyield a tan solid. Hydrolysis using KOH (2-5 eq) in 1:1 MeOH/H₂Oovernight would give the same result.

Preparation PP24

[1310] Resolution:

[1311] Purification of Partially Resolved Isoindoline-caboxylic acidmethyl ester: A solution of the crude material (97.62 g)isoindolinecaboxylic acid methyl ester in CH₂Cl₂ (350 mL) was extractedwith 1M HCl (400 mL, 200 mL). The combined aqueous portions were washedwith CH₂Cl₂ (4×250 mL) and then made basic with K₂CO₃ solution (85 g in150 mL of water). The mixture was extracted with CH₂Cl₂ (6×100 mL) andthe combined organic extracts were dried (Na₂SO₄) and concentrated togive partially resolved Isoindolinecaboxylic acid methyl ester as an oil(33.2 g). 60% ee by chiral CE.

Preparation PP25

[1312]

[1313] Resolution of Partially Resolved Isoindoline-caboxylic acidmethyl ester: A solution of partially resolved isoindoline-caboxylicacid methyl ester (33.24 g, 0.174 mol) in EtOH (130 mL) was treatedslowly with a solution of dibenzoyl-L-tartaric acid (56.06 g, 0.156 mol)in EtOH (200 mL). The solution was seeded with seeded with product andstirred at RT for 4 hours. Pure product was collected by filtration,washed with EtOH (30 mL) and dried to off-white crystals (60.49 g).96.5% ee by chiral CE.

Preparation PP26

[1314]

[1315] Resolution of N-BOC Isoindolinecaboxylic acid: A solution/slurryof racemic N-BOC Isoindolinecaboxylic acid (114.5 g, 0.413 mol) in EtOAc(1000 mL) was treated slowly with triethylamine (28.8 mL, 0.206 mol),followed by (S)-(−)-alpha-methylbenzylamine. The solution was seededwith product and stirred at RT overnight. The product was collected byfiltration, washed with EtOAc (200 mL) and dried to a white powder(62.98 g). 97.6% ee by chiral CE.

[1316] Asymmetric Hydrogenation Routes

[1317] Part I: Synthesis of the Z-isomer (precursor of asymmetrichydrogenation)

Preparation PP27

[1318] Z-isomer 5 was synthesized as outlined in Scheme P1. Compound 5was shown to be a single isomer by HPLC and H-1 nmr. The double bondstereochemistry was derived from comparative NOE data using thepurported E-isomer (Scheme P1). The best chiral induction was achievedusing compound 8/Ferrotane/MeOH-THF. With regard to the conversion of 9to 10, which would constitute a formal asymmetric synthesis ofisoindolene 10, this has been achieved using Super hydride-BF₃.OEt₂.However, the product was a mixture of 10 and the corresponding de-BOC(deprotected) compound.

Preparation PP28 Compound 2 (Scheme P1)

[1319] Phthalic anhydride (751.5 g, 5.014 mole), potassium acetate (498g, 5.014 mole) and acetic anhydride (1L) were stirred together undernitrogen. The mixture was slowly warmed to 145-150° C. and stirred for10 minutes, then at 140° C. for 20 minutes. The mixture was allowed toslowly cool to 80° C. over 1 hour. Three volumes of water were addedcausing precipitation of a solid. After filtration, the filtered solidwas washed with warm water and pulled as dry as possible for 30 minutes.The solid was then washed with ethanol and acetone respectively. Ifrequired further purification could be achieved by slurring the solid inacetone, at room temperature, for 15 minutes, then filtration. Drying invacuo at 50° C. for 20 hours gave compound 2 as an off-white solid, 470g (48%) with an NMR purity of approx. 90%.

Preparation PP29 Compound 3 (Scheme P1)

[1320] Compound 2 (470 g, 2.47 mole) was added to stirred aqueousammonia (470 ml conc. NH₃ in 4.7L water). The resultant mixture wasstirred at room temperature for 1 hour then filtered. The filtered solidwas washed with water. The combined aqueous filtrate and washings werecarefully acidified with 6M aq. HCl (2.35L). The precipitate was removedby filtration and dried in vacuo at 50° C. to give compound 3 as ayellow solid, 259 g (52%).

Preparation PP30 Compound 4 (Scheme P1)

[1321] Compound-3 (511 g, 2.7 mole) was slurried in toluene (10 vol).Thionyl chloride (385 g, 3.24 mole) was added over 10 minutes to thestirred mixture, which was then heated to reflux for 1.5 hours. H-1 NMRanalysis indicated approx. 80% conversion to acid chloride). DMF (3.7ml) was added and the mixture refluxed an additional 3 hours. Theresultant mixture was allowed to cool to 35° C. and methanol (1.27L)added at such a rate that the reaction temperature was maintained at30-35° C. The reaction mixture was kept at this temperature a further 15minutes then concentrated in vacuo to give compound 4 as a brown solid,536 g (quantitative).

Preparation PP31 Compound 5 (Scheme P1)

[1322] Compound 4 (750 g, 3.65 mole) was dissolved in acetonitrile(15L). The stirred mixture was cooled to 0-5° C. and DMAP (624 g, 5.11mole) added in one portion. After 10 minutes BOC anhydride (1115 g, 5.11mole) was added in one portion: there was a slight exotherm accompaniedby gas evolution. The mixture was stirred at room temperature for 5hours, and then concentrated in vacuo. The residue was dissolved inEtOAc and washed with 10% aqueous citric acid, satd. aq. Na₂CO₃ andwater respectively. After drying, concentration of the organics gave athick syrup. This material was run through a plug of silica gel (1.5 kg)eluting with 1:1 EtOAc-hexane. Compound 5 was isolated as a dark solid,619 g (55%). Careful chromatography on silica gel eluting with 20%EtOAc-hexane gave 5 as a fluffy white solid.

Scheme P2

[1323] Part II: Synthesis of the E-isomer (Precursor of asymmetrichydrogenation)

Preparation PP32

[1324] The E-isomer of Compound 8 (Scheme P2) was prepared as shown inScheme P2.

Preparation PP33 Compound 7 (Scheme P2)

[1325] The compound 7 was prepared according to the procedure of Einhornet al, Synth. Commun. 2001, 31(5), 741-748.

Preparation PP34 Compound 8 (Scheme P2)

[1326] Compound 7 (15.00 g, 60.7 mmole) and methyl(triphenylphosphoranylidene) acetate (41.40 g, 121.3 mmole) were slurred intoluene (150 ml). The mixture was stirred at reflux and monitored forreaction of 7 by GC. After 1.5 hours the reaction appears complete byGC. After cooling to room temperature, the mixture was filtered. Thesolid on the filter was washed with toluene until colorless. Thecombined filtrate/washings were concentrated in vacuo to leave a tansolid. This material was coated on silica gel and chromatographed onsilica gel (1 kg) eluting with 10% EtOAc-hexane. Compound 8 was isolatedas a white or pale yellow powder, 5.52 g (30%).

Preparation PP35

[1327] Screening of chiral hydrogenation conditions indicated that thebest chiral induction was achieved using compound 8/Ferrotane/MeOH-THF.With regard to the conversion of 9 to 10, which would constitute aformal asymmetric synthesis of isoindolene 10, this has been achievedusing Super hydride-BF₃.OEt₂. However, the product was a mixture of 10and the corresponding de-BOC (deprotected) compound.

Preparation PP36 Compound 15 (Scheme P4)

[1328] Tartrate salt 14 (58.00 g, 100.27 mmole) was slurried in water(580 ml). Solid NaHCO₃ (25.27 g, 300.8 mmole) was carefully added. BOCanhydride (22.98 g, 105.28 mmole) was added in one portion and theprogress of the reaction monitored by reverse phase HPLC. After 1 houradditional BOC anhydride (2.18 g, 10.00 mmole) was added. The reactionwas complete (by HPLC) after 3 hours. The mixture was extracted withEtOAc (2×250 ml). The combined organic extracts were washed with water(250 ml) and dried (MgSO₄). Filtration and concentration in vacuo gave15 as a clear light brown oil (31.33 g) contaminated with a small amountof t-BuOH and BOC anhydride. This material was used directly in the nextreaction.

Preparation PP37 Compound 16 (Scheme P4)

[1329] Ester 15 (29.21 g, 100.26 mmole) was dissolved in 3:1 THF-water(100 ml). LiOH (6.00 g, 250.65 mmole) was added in 1 portion to thestirred solution. After 17 hours, the mixture was stripped to drynessand the residue dissolved in water (500 ml). EtOAc (250 ml) was addedand solid NaHSO₄ added to the stirred mixture until the pH=3. Theorganic layer was separated and the aqueous layer extracted with EtOAc(250 ml). The combined EtOAc layers were dried (MgSO₄). Filtration andconcentration in vacuo gave acid 16 as a light tan solid, 27.10 g (97%).

[1330] The chemistry used is shown in Scheme P5. Two protocols wereused: method A used isolated 16, method B used a solution of 16 derivedfrom resolved salt 19.

Preparation PP38 Compound 17 (Scheme P5, method A)

[1331] Acid 16 (24.18 g, 87.2 mmole) and D-chloro-phenylalaninehydrochloride (21.81 g, 87.2 mmole) were dissolved in CH₂Cl₂ (100 ml)and DMF (25 ml). The mixture was stirred at ambient temperature. HOBT(13.55 g, 100.3 mmole) and Hunig's base (45.6 ml, 33.81 g, 261.6 mmole)were added. HATU (38.13 g, 100.3 mmole) was added in 1 portion (therewas a rapid exotherm to 50° C.). The mixture was stirred for 90 minutesthen diluted with EtOAc (750 ml). The resulting mixture was washed withwater, 5% KHSO₄, brine and satd. NaHCO₃ respectively, then dried.Filtration and concentration in vacuo gave crude 17 as a brown foam. Theproduct was purified by chromatography on silica gel (1 kg) eluting with1:1 EtOAc-hexane. Ester 17 was isolated as a tan powder, 38.85 g (94%).

Preparation PP39 Compound 17 (Scheme P5, Method B)

[1332] Resolved salt 19 (96.27 g, 232.5 mmole) was partitioned betweenwater (500 ml) and CH₂Cl₂ (250 ml) Solid KHSO₄ was added portion wiseuntil pH=2.5. Separate the organic layer and extract the aqueous layerwith CH₂Cl₂ (150 ml). The combined organic layers were dried (MgSO₄)then filtered. To this solution was added 4-chloro-D-phenylalanine(58.16 g, 232.5 mmole), HOBT (34.57 g, 255.8 mmole), Hunig's base (93.2ml, 69.13 g, 534.9 mmole) and finally HATU (97.26 g, 255.8 mmole). Theresultant mixture was stirred at room temperature for 18.5 hours, andthen poured onto a plug of silica gel (1 kg). This was washed with 1:1EtOAc-hexane until no more product elutes. Ester 17 was isolated as apink foam, 101.79 g (93%): contains about 1% unreacted 16.

Preparation PP40 Compound 18 (Scheme P5)

[1333] Ester 17 (38.64 g, 81.7 mmole) was dissolved in 3:1 THF-water(200 ml). LiOH (2.15 g, 89.9 mmole) was added to the mixture, which wasstirred at room temperature for 2 hours. The solvent was then removed invacuo and the residual solid taken up in water (600 ml). This wasextracted with MTBE (250 ml). The aqueous layer was separated andstirred with EtOAc (250 ml), and solid KHSO₄ was added portion wiseuntil pH=3. The layers were separated and the aqueous extracted withEtOAc (250 ml). The combined organic layers were dried over MgSO₄.Filtration and concentration in vacuo gave acid 18 as a light pink foam,38.41 g (35.71 g corrected for residual solvent, 95%).

[1334] In a 22 L 4-neck round bottom flask equipped with a refluxcondenser, thermocouple and nitrogen inlet, a slurry of 1000 g (5.4moles) of m-tyrosine in 10 L of 2B-3 EtOH was cooled to 5° C. To theslurry, 350 mL (12.4 moles) of thionyl chloride were added dropwise viaan addition funnel at such a rate to maintain the reaction temperaturebelow 20° C. Upon completion of addition, the reaction was heated toreflux temperature and stirred for 18 hrs. The reaction was concentratedto one-third the volume and 8 L of MTBE were charged. The resultingthick slurry was stirred for 14 hrs in a rotary evaporator at ρ.τ. Theresulting solid was isolated on a filter pad and dried at 40° C. for 48hrs yielding 1288 g (95%). NMR (DMSOd6) indicated desired material.

Preparation PP42

[1335]

[1336] In a 22L 4 neck round bottom flask equipped with a mechanicalstirrer, thermocouple, and reflux condenser placed on top of a Soxhletextractor charged with 4° A sieves, a semi-solution of m-tyrosine ethylester hydrochloride 1288 g (5.26 moles) in 13 L of acetone was heated toreflux temperature. The condensate was filtered through the sieves toremove water. The reaction was stirred vigorously at reflux for 48 hrs.An NMR sample in DMSOd₆ indicated the absence of starting material. Thereaction was cooled to rt and concentrated to yield an off-white solid,1411 g (94%).

Preparation PP43

[1337] Step 3: Triflation

[1338] In a 22 L 4 neck round bottom flask equipped with a refluxcondenser, mechanical stirrer, nitrogen inlet, and a thermocouple, 1240g (4.35 moles) of the starting material salt in 12.4 L of methylenechloride was cooled to 4° C. To the mixture, 1452 mL (10.4 moles) oftriethylamine were added and stirred into solution. Triflic anhydride,1472 mL (5.22 moles) was added dropwise to the reaction at such a rateto maintain the internal temperature below 10° C. The ice bath wasremoved and the reaction warmed to ρ.τ. and stirred for 18 hrs. Thereaction was concentrated to a oil then dissolved in 4 L of EtOAc andconcentrated again to an oil in an effort to remove excess triflicanhydride The crude residue was dissolved in 4 L of EtOAc and washedwith water and saturated sodium bicarbonate solution. The organic layerwas isolated and dried with sodium sulfate, filtered and concentrated toyield 1720 g (>100%) of a crude dark oil which was used without furtherpurification.

Preparation PP44

[1339] Step 4: Deoxygenation

[1340] A solution of 1720 g (4.35 moles) of crude starting material in14 L of acetone was charged to a 10 gallon stainless steel autoclave. Tothe solution, a slurry of 5% Pd/C in 1.2 L of toluene was added. Thereaction mixture was evacuated and purged with H₂ gas at 50 psi twotimes. The reaction was stirred overnight at 50° C. with H₂ at 50 psi. Asample aliquot indicated no reaction had occurred. The mixture wasfiltered and concentrated to a thick oil and resubjected to reactionconditions. After 18 hrs, NMR of a sample aliquot indicated absence ofstarting material. The reaction mixture was filtered and the filtrateconcentrated to yield 1581 g of an off-white solid (95%).

Preparation PP45

[1341] Step 5: Hydrolysis/Salt Formation

[1342] To a 2 L 3 neck round bottom flask equipped with a mechanicalstirrer, thermocouple, and nitrogen inlet, a mixture of 700 g (1.83moles) of the triflate salt starting material was charged. A solution of427 g. (1.83 moles) of the starting material free base in 13.3 L of THFwas added followed by 700 mL of water. The semi-solution was stirredvigorously at r.t. To the reaction flask, 43.7 g (1.83 moles) of solidLiOH were added in small portions at such a rate to maintain theinternal temperature below 35° C. The reaction was stirred for 18 hrs atr.t and concentrated to yield a thick oil. THF (4 L) was added and thesemi-solution was concentrated. This was repeated with toluene and thesemi-solid was placed under house vacuum on the roto vap with stirringfor 18 hrs to yield 650 g of a crude solid. The solid was reslurred inEtOAc, filtered, isolated and dried to yield 525 g (68%) of the lithiumsalt as an off-white solid.

Preparation PP46

[1343] Step 6: Coupling

[1344] Solid d-chloro-phenylalanine 446 g (1.78 moles) was added to thesemi-solution followed by 20 g (0.162 moles) of DMAP. The resultingmixture was stirred for 15 minutes then solid EDCl(1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) 390 g(2.03 moles) was added. The reaction mixture was heated to 80° C. andstirred for 18 hours. Thin layer chromatography (1:1 EtOAc:Hex)indicated very little starting material present. The reaction was cooledto rt and concentrated to yield a thick oil. The crude oil was dissolvedin EtOAc and washed with water, and brine. The solution was dried withsodium sulfate, filtered and concentrated to yield a thick oil, 426 g.The crude oil was chromatographed in several lots using a Waters Prep500 chromatography apparatus. The eluent consisted of a gradient system,5%-80% EtOAc in heptane at a flow rate of 240 ml/min over 38 minutes.The two diasteromers were separated and isolated to yield 119.04 g forthe top spot and 111.3 g for the bottom spot. Conformation of bothdesired diastereomers was achieved via NMR (DMSO₆).

Preparation PP47

[1345] Resolution of tetrahydroisoquinolinecarboxylic acid ethyl esterto prepare 1-tartaric acid salt:

[1346] Preparation of free-base: A racemic mixture oftetrahydroisoquinolinecarboxylic acid (7.43 g) in EtOAc (60 mL) wastreated with saturated NaHCO₃ solution (60 mL) and saturated Na₂CO₃solution (10 mL). The mixture was agitated and the layers wereseparated. The organic phase was dried (Na₂SO₄) and concentrated to givethe corresponding free-base as an oil (4.85 g)

[1347] Resolution: A mixture of the above free base (467 mg, 2.0 mmol),and L-tartaric acid (300 mg, 2.0 mmol) in acetone (4 mL) was stirred atRT overnight. The title L-tartaric acid salt was collected byfiltration, washed with acetone (about 2 mL) and dried to a white powder(367 mg). 100% ee by chiral CE.

Preparation PP48

[1348] Resolution of N-BOC tetrahydroisoquinolinecarboxylic acid

[1349]2-{2-[(tert-butyl)oxycarbonyl]-1,2,3,4-tetrahydro-isoquinolyl}aceticacid dehydroabietylamine salt: Racemic2-{2-[(tert-butyl)oxycarbonyl]-1,2,3,4-tetrahydroisoquinolyl}acetic acid(30.15 g, 103.5 mmol) was dissolved in i-PA (300 mL).Dehydroabietylamine (22.11 g, 52.7 mmol of a 68 weight % mixture) wasadded to the solution, which was then agitated on a multi-arm shaker for63 h. The resultant thick paste was filtered and rinsed with i-PA (50mL, 25 mL). Dried in a 50° C. vacuum oven to obtain a white solid (27.73g, 52% ee by chiral CE analysis). The product was reslurried in i-PA(266 mL) and agitated on a multi-arm shaker for 23.5 h. Filtered thethick slurry and rinsed with cold i-PA (50 mL, 30 mL). Dried the cake ina 50° C. vacuum oven and obtained the product as a white solid (23.63 g,40% yield, 94% ee by chiral CE analysis).

Preparation PP49

[1350] Enamine 21 (Scheme P6) was prepared as a substrate for asymmetrichydrogenation screening studies. It is formed as an approx. 10:1 mixturewith imine 22. The enamine (21) may be NH-protected i.e., by a Bocprotecting group. The resulting compound 23 may be subjected toasymmetric hydrogenation to afford the acetic acid or methylacetatesubstituted isoquinoline, which may be processed into a compound offormula I as demonstrated previously.

Preparation PP50 Compound 21 (Scheme P6)

[1351] Prepared as published W Sobotka et al, J. Org. Chem., 1965, 30,3667

Preparation PP51

[1352] The chiral synthesis of gem-dimethyl TIC using L-Dopa as thestarting material instead of tyrosine was successfully demonstrated upto the Pictet-Spengler reaction with L-DOPA and acetone. The product isa mixture of starting material 24 and product 25 (major component). Theproduct was isolated by using common isolation procedures. Analternative isolation method is to react the mixture (24 and 25) withBOC anhydride wherein the less hindered N—H in 24 leads to preferentialBOC protection of 24, allowing for ready separation of 25. Chemistry forthe rest of the sequence i.e., deoxygenation reaction, has beendemonstrated herein.

What is claimed is:
 1. A compound of formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, whereinL and L¹ are independently: hydrogen or together oxo; T is:

R is independently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl,C₁-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂₎ _(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, OD)[O]_(q)(CH₂)_(n) heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; R¹ is independently: hydrogen, CONH(C₁-C₈ alkyl),C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇ cycloalkyl or oxo, provided that oxo isnot attached to the same carbon that attached to nitrogen which forms anamide bond; R³ is independently: aryl or thienyl; wherein aryl andthienyl are optionally substituted with one to three substituentsselected from the group consisting of: cyano, halo, C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy, C₁-C₄ haloalkyl and C₁-C₄haloalkyloxy; R⁴ is independently: hydrogen, C₁-C₈ alkyl, C(O)R⁹,C(O)OR⁹, C₃-C₇ cycloalkyl or (CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂ C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy, heteroaryl, hydroxy,heterocyclyl, wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹ (D)OR⁹, (D)OCOR⁹,(D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl, heteroaryl,heterocyclyl, alkyl or cycloalkyl is optionally substituted with one tothree substituents selected from the group consisting of oxo, C₁-C₈alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ is independently:hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇ cycloalkyl, (D)aryl,wherein aryl being phenyl or naphthyl, (D)heteroaryl or (D)heterocyclyl;wherein heterocyclyl excludes a heterocyclyl containing a singlenitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl; each R¹¹ is independently:hydrogen, C₁-C₈ alkyl, (D)aryl, (D)heteroaryl (CH₂)_(n)N(R⁸)₂,(CH₂)_(n)NR⁸C(O)C₁-C₄ alkyl, (CH₂)_(n)NR⁸SO₂C₁-C₄ alkyl,(CH₂)_(n)SO₂N(R⁸)₂, (CH₂)_(n)[O]_(q)C₁-C₈ alkyl,(CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸COR⁸, (CH₂)_(n)[O]_(q)(CH₂)_(n)NR⁸SO₂R⁸,(CH₂)_(n)[O]_(q)-heterocyclyl or (CH₂)_(n)[O]_(q)(C₁-C₈alkyl)-heterocyclyl; and wherein n is 2-8; each R¹² is independently:hydrogen, C₁-C₈ alkyl, (D)phenyl C(O)C₁-C₈ alkyl, C(O)phenyl, SO₂C₁-C₈alkyl or SO₂-phenyl; D is a bond or —(CH₂)_(n)—; n is 0-8; p is 0-5; qis 0-1; and r is 1-2.
 2. The compound of claim 1, wherein R³ is phenyloptionally para-substituted with chloro, bromo, fluoro, iodo, methoxy,benzyloxy or methyl.
 3. The compound of claim 2, wherein R³ is phenylpara-substituted with chloro, fluoro or methoxy.
 4. The compound ofclaim 3, wherein R⁴ is hydrogen.
 5. The compound of claim 4, wherein—(CH₂)_(n)—T is:

where * denotes a chiral carbon atom having a R or S configuration. 6.The compound of claim 5, wherein L and L¹ are together oxo and thechiral carbon has R configuration.
 7. A compound of formula II,

or a pharmaceutically acceptable salts or stereoisomers thereof, whereinP is 0-5; n is 0-8; q is 0-1; D is a bond or —(CH₂)_(n)—; R isindependently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl, C₁-C₈alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n)heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy,heteroaryl, hydroxy, heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹(D)OR⁹, (D)OCOR⁹, (D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl,heteroaryl, heterocyclyl, alkyl or cycloalkyl is optionally substitutedwith one to three substituents selected from the group consisting ofoxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ isindependently: hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇cycloalkyl, (D)aryl, wherein aryl being phenyl or naphthyl heteroaryl orheterocyclyl; wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; and each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl.
 8. A compound of formula III,

or a pharmaceutically acceptable salts or stereoisomers thereof, whereinP is 0-5; n is 0-8; q is 0-1; D is a bond or —(CH₂)_(n)—; R isindependently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl,C_(l)-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n)heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy,heteroaryl, hydroxy, heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹(D)OR⁹, (D)OCOR⁹, (D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl,heteroaryl, heterocyclyl, alkyl or cycloalkyl is optionally substitutedwith one to three substituents selected from the group consisting ofoxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ isindependently: hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇cycloalkyl, (D)aryl, wherein aryl being phenyl or naphthyl heteroaryl orheterocyclyl; wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; and each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl.
 9. A compound of formula IV,

or a pharmaceutically acceptable salts or stereoisomers thereof, whereinP is 0-5; n is 0-8; q is 0-1; D is a bond or —(CH₂)_(n)—; R isindependently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl,C_(l)-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n) heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy,heteroaryl, hydroxy, heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹(D)OR⁹, (D)OCOR⁹, (D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl,heteroaryl, heterocyclyl, alkyl or cycloalkyl is optionally substitutedwith one to three substituents selected from the group consisting ofoxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ isindependently: hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇cycloalkyl, (D)aryl, wherein aryl being phenyl or naphthyl heteroaryl orheterocyclyl; wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; and each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl.
 10. The compound of claim 9,wherein R¹⁰ is hydrogen or (C₁-C₈)alkyl.
 11. A compound of formula V,

or a pharmaceutically acceptable salts or stereoisomers thereof, whereinP is 0-5; n is 0-8; q is 0-1; D is a bond or —(CH₂)_(n)—; R isindependently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C-₈ alkyl,C_(l)-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n) heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂C₁-C₈ alkyl, (D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy,heteroaryl, hydroxy, heterocyclyl, wherein heterocyclyl excludes aheterocyclyl containing a single nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹(D)OR⁹, (D)OCOR⁹, (D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl,heteroaryl, heterocyclyl, alkyl or cycloalkyl is optionally substitutedwith one to three substituents selected from the group consisting ofoxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ isindependently: hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇cycloalkyl, (D)aryl, wherein aryl being phenyl or naphthyl heteroaryl orheterocyclyl; wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; and each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl.
 12. A compound selected fromthe group consisting of: Name of Compounds Structure of Compounds1-(D-Tic-4-Cl-D-Phe)-4-(2- methanesulfonylamino-phenyl)- piperazine

1-(D-Tic-4-Cl-D-Phe)-4-(2- dimethylaminomethyl-phenyl)- piperazine

3-(4-chloro-phenyl)-2-[(1,2,3,4- tetrahydro-isoquinolin-3-ylmethyl)-amino]-1-[4-(2-[1,2,4]triazol-1- ylmethyl-phenyl)-piperazin-1-yl]-propan-1-one

N-(1-(4-chloro-benzyl)-2-{4-[2- (isobutyl-methanesulfonyl-amino)-phenyl]-piperazin-1-yl}-2-oxo-ethyl)-2- (2,3-dihydro-1H-isoindol-1-yl)-acetamide


13. A pharmaceutical composition which comprises a pharmaceuticalcarrier and at least one compound of formula I or its pharmaceuticallyacceptable salts or stereoisomers thereof as recited in claim
 1. 14. Thepharmaceutical composition of claim 13, which comprises a second activeingredient selected from the group consisting of: an insulin sensitizer,insulin mimetic, sulfonylurea, alpha-glucosidase inhibitor, HMG-CoAreductase inhibitor, sequestrant cholesterol lowering agent, beta 3adrenergic receptor agonist, neuropeptide Y antagonist, phosphodiester Vinhibitor, and an alpha 2 adrenergic receptor antagonist.
 15. A processof making a pharmaceutical composition comprising a compound of formulaI or its pharmaceutically acceptable salt or stereoisomers thereof asrecited in claim 1 and a pharmaceutically acceptable carrier.
 16. Amethod of preventing or treating obesity in a mammal comprising theadministration of a therapeutically effective amount of the compound offormula I as recited in claim
 1. 17. A method of preventing or treatingdiabetes mellitus in a mammal comprising the administration of atherapeutically effective amount of the compound of formula I as recitedin claim
 1. 18. A method of preventing or treating male or female sexualdysfunction in a mammal comprising the administration of atherapeutically effective amount of the compound of formula I as recitedin claim
 1. 19. The method of 18, wherein the male or female sexualdysfunction is erectile dysfunction.
 20. A process for preparing acompound of formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein—CLL¹—(CH₂)_(n)—T is:

wherein R₁ is hydrogen, C₁-C₈ alkyl, Boc, CBZ, phenyl, FMOC or (C₁-C₈alkyl)phenyl; Q represents a moiety:

R is independently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl,C₁-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n) heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; R¹ is independently: hydrogen, CONH(C₁-C₈ alkyl),C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇ cycloalkyl or oxo, provided that oxo isnot attached to the same carbon that attached to nitrogen which forms anamide bond; R³ is independently: aryl or thienyl; wherein aryl andthienyl are optionally substituted with one to three substituentsselected from the group consisting of: cyano, halo, C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy, C₁-C₄ haloalkyl and C₁-C₄haloalkyloxy; R⁴ is independently: hydrogen, C₁-C₈ alkyl, C(O)R⁹,C(O)OR⁹, C₃-C₇ cycloalkyl or (CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂ C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy, heteroaryl, hydroxy,heterocyclyl, wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹ (D)OR⁹, (D)OCOR⁹,(D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl, heteroaryl,heterocyclyl, alkyl or cycloalkyl is optionally substituted with one tothree substituents selected from the group consisting of oxo, C₁-C₈alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ is independently:hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇ cycloalkyl, (D)aryl,wherein aryl being phenyl or naphthyl, (D)heteroaryl or (D)heterocyclyl;wherein heterocyclyl excludes a heterocyclyl containing a singlenitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl; D is a bond or —(CH₂)_(n)—; nis 0-8; p is 0-5; q is 0-1; and r is 1-2; comprising the steps of: a)reacting a compound having a structural formula 1,

 with CH₂CH═C(O)OR^(a) wherein R^(a) is hydrogen or C₁-C₈ alkyl and X ishalo, in the presence of a catalyst and a base in a suitable organicsolvent to give the compound of formula 2,

b) reductively aminating the compound of formula 2 in the presence ofamine in an acidic condition to give a compound of formula 3,

c) cyclizing the compound of formula 3 by Michael addition to give acompound of formula 4 or stereoisomers thereof,

d) coupling the compound of formula 4 or stereoisomers thereof, whereinR^(a) of compound 4 is H, with a compound of formula 5,

 wherein R^(a) of compound 5 is C₁-C₈ alkyl, to give a compound offormula 6;

e) coupling the compound of formula 6, wherein R^(a) is H, with acompound having a structural,

 to afford the compound of formula
 1. 21. The process of claim 20,wherein

in Step (a) is 2-boromobenzaldehydes.
 22. The process of claim 21,wherein CH₂CH═C(O)OR in Step (a) is methylacrylate.
 23. The process ofclaim 22, wherein the catalyst in Step (a) is selected from the groupconsisting of: Pd(Ph₃P)₂Cl₂, Pd(Ph₃P)₄Cl₂, Pd(Ph₃P)₄, Pd(Ph₃P)₂Cl₂/CuI,Pd(OAc)₂/Ph₃P-Bu₄NBr, Pd(Ph₃P)₄Cl₂/H₂ and Pd(OAc)₂/P(O-tol)₃; andwherein the base in Step (a) is NR₃ wherein R is hydrogen or C₁-C₈alkyl.
 24. The process of claim 23, wherein the amine in Step (b) isselected from the group consisting of: benzylamine,alpha-methylbenzylamine and BocNH₂.
 25. The process of claim 24, whereinthe Step (b) further comprises reducing of intermediate imine compoundin the presence of reducing agent, the reducing agent being selectedfrom the group consisting of: NaCNBH₃, Na(OAc)₃BH, NaBH₄/H+, and acombination of Et₃SiH and TFA in CH₃CN or CH₂Cl₂.
 26. The process ofclaim 25, wherein the stereoisomer of compound of formula 4 in Step (c)is a compound of formula 4a.


27. The process of claim 26, wherein the compound of formula 4a isprepared by asymmetric hydrogenation of a compound having structuralformula,


28. The process of claim 27, wherein the Michael addition in Step (c) iscarried out in a basic workup condition.
 29. The process of claim 20,wherein the Step (e) further comprises deprotecting or protecting of thecompound of formula (4) at NR₁.
 30. A process for preparing a compoundof formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein—CLL¹—(CH₂)_(n)—T is:

Q represents a moiety:

R is independently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl,C₁-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n)heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; R¹ is independently: hydrogen, CONH(C₁-C₈ alkyl),C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇ cycloalkyl or oxo, provided that oxo isnot attached to the same carbon that attached to nitrogen which forms anamide bond; R³ is independently: aryl or thienyl; wherein aryl andthienyl are optionally substituted with one to three substituentsselected from the group consisting of: cyano, halo, C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy, C₁-C₄ haloalkyl and C₁-C₄haloalkyloxy; R⁴ is independently: hydrogen, C₁-C₈ alkyl, C(O)R⁹,C(O)OR⁹, C₃-C₇ cycloalkyl or (CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂ C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy, heteroaryl, hydroxy,heterocyclyl, wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹ (D)OR⁹, (D)OCOR⁹,(D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl, heteroaryl,heterocyclyl, alky or cycloalkyl is optionally substituted with one tothree substituents selected from the group consisting of oxo, C₁-C₈alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ is independently:hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇ cycloalkyl, (D)aryl,wherein aryl being phenyl or naphthyl, (D)heteroaryl or (D)heterocyclyl;wherein heterocyclyl excludes a heterocyclyl containing a singlenitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl; each R¹¹ is independently:hydrogen or (C₁-C₈)alkyl; D is a bond or —(CH₂)_(n)—; n is 0-8; p is0-5; q is 0-1; and r is 1-2; comprising the steps of: a) esterifying acompound of formula 1,

 with an alcohol R^(a)OH to form a compound of formula 2,

 wherein R^(a) is C₁-C₄ alkyl or (D)phenyl; b) reacting a compound offormula 2 with R¹¹COR¹¹ to form a compound of formula 3,

 wherein R¹¹ is independently hydrogen or C₁-C₄ alkyl; c) reacting acompound of formula 3 with an activating group to form a compound offormula 4,

 wherein A is an activating group; d) deoxygenating the compound offormula 4 by hydrogenation to afford a compound of formula 5,

e) optionally reacting the compound of formula 5 with an inorganic baseto form a compound of formula 6,

 wherein HA is an acidic and M is a univalent cation; f) resolving thecompound of formula 5 or formula 6 to afford a chiral compound offormula 7,

 wherein M is hydrogen and R^(a′) is H or R^(a); g) coupling thecompound of formula 7 with a compound of formula 8,

 to afford a compound of formula 9,

h) coupling the compound of formula 9 with a compound having a formula,

 to afford a compound of formula I.
 31. A process for preparing acompound of formula I:

or a pharmaceutically acceptable salts or stereoisomers thereof, wherein—CLL¹—(CH₂)_(n)—T is:

Q represents a moiety:

R is independently: hydrogen, hydroxy, cyano, nitro, halo, C₁-C₈ alkyl,C₁-C₈ alkoxy, C₁-C₄ haloalkyl, (D)C(O)R⁹, (D)C(O)OR⁹, (D)C(O)SR⁹,(D)C(O)heteroaryl, (D)C(O)heterocyclyl, (D)C(O)N(R⁹)₂, (D)N(R⁹)₂,(D)NR⁹COR⁹, (D)NR⁹CON(R⁹)₂, (D)NR⁹C(O)OR⁹, (D)NR⁹C(R⁹)═N(R⁹),(D)NR⁹C(═NR⁹)N(R⁹)₂, (D)NR⁹SO₂R⁹, (D)NR⁹SO₂N(R⁹)₂,(D)NR⁹(CH₂)_(n)heterocyclyl, (D)NR⁹(CH₂)_(n)heteroaryl, (D)OR⁹, OSO₂R⁹,(D)[O]_(q)(C₃-C₇ cycloalkyl), (D)[O]_(q)(CH₂)_(n)aryl,(D)[O]_(q)(CH₂)_(n)heteroaryl, (D)[O]_(q)(CH₂)_(n) heterocyclyl, whereinheterocyclyl excludes a heterocyclyl containing a single nitrogen whenq=1, (D)SR⁹, (D)SOR⁹, (D)SO₂R⁹, or (D)SO₂N(R⁹)₂; wherein C₁-C₈ alkyl,C₁-C₈ alkoxy, C₃-C₇ cycloalkyl, aryl, heterocyclyl and heteroaryl areoptionally substituted with one to five substituents independentlyselected from R⁸; R¹ is independently: hydrogen, CONH(C₁-C₈ alkyl),C₁-C₈ alkyl, (D)phenyl, (D)C₃-C₇ cycloalkyl or oxo, provided that oxo isnot attached to the same carbon that attached to nitrogen which forms anamide bond; R³ is independently: aryl or thienyl; wherein aryl andthienyl are optionally substituted with one to three substituentsselected from the group consisting of: cyano, halo, C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ alkoxy, C₁-C₄ haloalkyl and C₁-C₄haloalkyloxy; R⁴ is independently: hydrogen, C₁-C₈ alkyl, C(O)R⁹,C(O)OR⁹, C₃-C₇ cycloalkyl or (CH₂)_(n)O(C₁-C₈ alkyl), wherein n is 2-8;each R⁸ is independently: hydrogen, halo, oxo N(R¹⁰)₂ C₁-C₈ alkyl,(D)C₃-C₇ cycloalkyl, C₁-C₄ haloalkyl, C₁-C₄ alkoxy, heteroaryl, hydroxy,heterocyclyl, wherein heterocyclyl excludes a heterocyclyl containing asingle nitrogen, phenyl, (D)COR⁹, (D)C(O)OR⁹ (D)OR⁹, (D)OCOR⁹,(D)OCO₂R⁹, (D)SR⁹, (D)SOR⁹, or (D)SO₂R⁹; wherein aryl, heteroaryl,heterocyclyl, alkyl or cycloalkyl is optionally, substituted with one tothree substituents selected from the group consisting of oxo, C₁-C₈alkyl, N(R¹⁰)₂, OR¹⁰, SR¹⁰ and CO₂R¹⁰; each R⁹ is independendy:hydrogen, C₁-C₈ alkyl, C₁-C₄ haloalkyl, (D)C₃-C₇ cycloalkyl, (D)aryl,wherein aryl being phenyl or naphthyl, (D)heteroaryl or (D)heterocyclyl;wherein heterocyclyl excludes a heterocyclyl containing a singlenitrogen; and wherein aryl, heteroaryl, heterocyclyl, alkyl orcycloalkyl is optionally substituted with one to three substituentsselected from the group consisting of oxo, C₁-C₈ alkyl, N(R¹⁰)₂, OR¹⁰,SR¹⁰ and CO₂R¹⁰; each R¹⁰ is independently: hydrogen, (C₁-C₈)alkyl,C(O)C₁-C₈ alkyl, aryl or C₃-C₇ cycloalkyl; each R¹⁰ is independendy:hydrogen or (C₁-C₈)alkyl; D is a bond or —(CH₂)_(n)—; n is 0-8; p is0-5; q is 0-1; and r is 1-2; comprising the steps of: a) reacting acompound formula 1:

 wherein X is halo, and R¹¹ is independently, hydrogen or C₁-C₄ alkyl,with CNCH₂CO₂R^(a) wherein R^(a) is C₁-C₈ alkyl or benzyl to afford acompound of formula 2:

b) protecting the compound of formula 2 to form the compound of formula3:

c) hydrogenating the compound of formula 3 to afford a compound offormula 4:

d) coupling the compound of formula 4 wherein R^(a′) is hydrogen orR^(a) with a compound of formula 5,

 to afford a compound of formula 6,

e) coupling the compound of formula 6 with a compound having a formula,

 to afford a compound of formula I.